WO2008040002A2 - Procédés, compositions et articles de fabrication de composés modulant le hif - Google Patents

Procédés, compositions et articles de fabrication de composés modulant le hif Download PDF

Info

Publication number
WO2008040002A2
WO2008040002A2 PCT/US2007/079948 US2007079948W WO2008040002A2 WO 2008040002 A2 WO2008040002 A2 WO 2008040002A2 US 2007079948 W US2007079948 W US 2007079948W WO 2008040002 A2 WO2008040002 A2 WO 2008040002A2
Authority
WO
WIPO (PCT)
Prior art keywords
compound
formula
biological matter
organism
stasis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2007/079948
Other languages
English (en)
Other versions
WO2008040002A9 (fr
WO2008040002A3 (fr
Inventor
Mark B. Roth
Mark Budde
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fred Hutchinson Cancer Center
Original Assignee
Fred Hutchinson Cancer Center
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fred Hutchinson Cancer Center filed Critical Fred Hutchinson Cancer Center
Publication of WO2008040002A2 publication Critical patent/WO2008040002A2/fr
Anticipated expiration legal-status Critical
Publication of WO2008040002A9 publication Critical patent/WO2008040002A9/fr
Publication of WO2008040002A3 publication Critical patent/WO2008040002A3/fr
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/12—Chemical aspects of preservation
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/12—Chemical aspects of preservation
    • A01N1/122—Preservation or perfusion media
    • A01N1/126—Physiologically active agents, e.g. antioxidants or nutrients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/095—Sulfur, selenium, or tellurium compounds, e.g. thiols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • the present invention relates generally to the field of cell biology and physiology. More particularly, it concerns methods, compositions and apparatuses for enhancing survivability of and/or reducing damage to cells, tissues, organs, and organisms, particularly under adverse conditions, including but not limited to hypoxic or ischemic states, using one or more substances, including those that compete with oxygen.
  • the present invention includes methods, compositions and apparatuses for treating, preventing, and diagnosing diseases and conditions by exposing a subject to a compound, such as an oxygen antagonist, protective metabolic agent, or other chemical compound discussed herein, or a precursor thereof, that can achieve its stated goal (collectively referred to as
  • compositions and apparatuses of the present invention relate to the modulation of the ⁇ -subunt of hypoxia inducible factor (HIF), an oxygen-responsive transcriptional activator.
  • HIF hypoxia inducible factor
  • hypoxia and ischemia are two physiological states characterized by reductions in oxygen and blood flow, respectively.
  • hypoxic and ischemic states are unfavorable, and treatments exist to combat these states and their related conditions, such as anemia, myocardial infarction, stroke and occlusive arterial disease.
  • hypoxic and/or ischemic states may be preferable, such as during organ transplantation or prior to the inducement of trauma.
  • ischemic and hypoxic disorders are focused on relief of symptoms and treatment of causative disorders.
  • treatments for myocardial infarction include nitroglycerin and analgesics to control pain and relieve the workload of the heart.
  • Other medications including digoxin, diuretics, amrinone, ⁇ -blockers, lipid-lowering agents and angiotensin-converting enzyme inhibitors, are used to stabilize the condition, but none of these therapies directly address the tissue damage produced by the ischemia and hypoxia.
  • hypoxia- and ischemia-related conditions Due to deficiencies in current treatments, there remains a need for compounds that are effective in treating hypoxia- and ischemia-related conditions.
  • hypoxia-related conditions such as anemia, including anemia associated with diabetes, ulcers, kidney failure, cancer, infection, dialysis, surgery, and chemotherapy.
  • Other conditions involving ischemia and hypoxia include, for example, occlusive arterial disease, angina pectoris, intestinal infarctions, pulmonary infarctions, cerebral ischemia, and myocardial infarction.
  • hypoxia-related conditions such as anemia, including anemia associated with diabetes, ulcers, kidney failure, cancer, infection, dialysis, surgery, and chemotherapy.
  • Other conditions involving ischemia and hypoxia include, for example, occlusive arterial disease, angina pectoris, intestinal infarctions, pulmonary infarctions, cerebral ischemia, and myocardial infarction.
  • hypoxia inducible factor a protein comprised of two subunits, the ⁇ -subunit of which is oxygen-regulated.
  • HIF ⁇ hypoxia inducible factor
  • hypoxia inducible factor a protein comprised of two subunits, the ⁇ -subunit of which is oxygen-regulated.
  • HIF ⁇ is hydroxylated and degraded but under hypoxic conditions, HIF ⁇ is not degraded and forms a complex with HIF ⁇ .
  • HIF ⁇ / ⁇ then regulates the expression of certain genes, such as erythropoeitin (EPO).
  • HIF ⁇ Since the accumulation of HIF ⁇ is associated with hypoxic and ischemic conditions, HIF ⁇ presents itself as a viable target for treatment of deleterious hypoxic and ischemic conditions as well as for the intentional inducement of such conditions that may be beneficial for a tissue, organ or organism. Indeed, compounds have been found that can modulate these conditions via HIF regulation. See, e.g., U.S. Pat. Appln. 2003/0176317; U.S. Pat. Appln. 2004/0254215, each of which is incorporated herein by reference in its entirety. Stasis is another physiological state that may be associated with reductions in oxygen and blood flow.
  • Stasis a cell, tissue or organ, or organism (collectively referred to as “biological material”) is living, but cellular functions necessary for cell division, developmental progression, and/or metabolic state are slowed or even stopped. This state is desirable in a number of contexts. Stasis can be used as a method of preservation by itself, or it may be induced as part of a cryopreservation regimen. For example, biological materials may be preserved for research use, for transportation, for transplantation, for therapeutic treatment (such as ex vivo therapy), and to prevent the onset of trauma.
  • tissue culture cells are often stored for periods of time in tanks that hold liquid nitrogen; however, these tanks frequently require that the liquid nitrogen in the unit be periodically replaced, otherwise it becomes depleted and the temperature is not maintained. Furthermore, damage to cells and tissue occurs as a result of the freeze/thaw process. Thus, improved techniques are needed. Stasis with respect to entire organisms has similar uses. For instance, transportation of organisms could be facilitated if they had entered stasis. This might reduce physical and physiological damage to the organism by reducing or eliminating stress or physical injury. Stasis may be beneficial by decreasing the need of the biological material for oxygen and, therefore, bloodflow.
  • hypoxia As discussed, the conditions of hypoxia, ischemia and stasis all involve reductions in physiological aspects such as oxygen, blood flow and metabolic rates.
  • the present invention relates to the interrelationship among hypoxic, ischemic and stasis conditions, based on the common feature of cellular HIF modulation by compounds and methods of the present invention.
  • the invention is generally drawn to compounds and methods that modulate HIF activity, such as by stabilizing HIF via the inhibition of HIF hydroxylation and thereby, in some embodiments, activating HIF-regulated gene expression.
  • the methods can be applied to the prevention, pretreatment, and/or treatment of conditions or states associated with HIF, such as stasis and anemic, ischemic and hypoxic conditions. These methods can be applied in vivo or in vitro.
  • HIF hypoxia inducible factor
  • the transcription factor HIF (hypoxia inducible factor) system is a key regulator of responses to hypoxia, occupying a central position in oxygen homeostasis in a wide range of organisms.
  • a large number of transcriptional targets have been identified, with critical roles in angiogenesis, erythropoiesis, energy metabolism, inflammation, vasomotor function, and apoptotic/proliferative responses.
  • the system is essential for normal development, and plays a key role in pathophysiological responses to ischaemia/hypoxia.
  • HIF is also important in cancer, in which it is commonly upregulated, and has major effects on tumor growth and angiogenesis.
  • the HIF DNA binding complex consists of a heterodimer of ⁇ and ⁇ subunits.
  • Regulation by oxygen occurs through hydroxylation of the ⁇ -subunits, typically of one or more proline residues, which are then rapidly destroyed by the proteasome in oxygenated cells.
  • pVHL or VHL von Hippel-Lindau tumor suppressor protein
  • the present invention relates generally to methods of stabilizing the ⁇ -subunit of HIF.
  • the HIF ⁇ is selected from the group consisting of HIF- l ⁇ , HIF-2 ⁇ , HIF-3 ⁇ and any fragment thereof.
  • the present invention provides, in one aspect, methods for stabilizing endogenous HIF ⁇ .
  • the HIF ⁇ is endogenous to the subject.
  • stabilization occurs via inhibition of HIF prolyl hydroxlation.
  • Embodiments of the present invention include methods for stabilizing HIF ⁇ in which a compound that stabilizes HIF ⁇ is administered to a subject in vivo.
  • the subject can be, for example, an animal, such as a mammal, wherein the mammal may be, for example, a human.
  • Methods of ex vivo administration are also contemplated.
  • the subject can be, e.g., a cell, tissue, or organ, etc.
  • the subject is a cell, tissue, or organ derived from a system such as the renal, cardiac, hepatic, pulmonary, hematopoietic, gastrointestinal, neuronal, or musculoskeletal system, etc.
  • the present invention provides a method for treating, preventing, or pretreating a HIF-associated condition, the method comprising stabilizing HIF ⁇ .
  • the invention provides a method for treatment, prevention, or pretreatment/preconditioning of a HIF-associated condition in a subject, the method comprising stabilization of HIF ⁇ .
  • the HIF- associated condition is associated with ischemia or hypoxia.
  • the HIF- associated condition is associated with stasis.
  • the method comprises administering to the subject an Effective Compound, as described herein, that stabilizes HIF ⁇ . Methods of administration of one or more Effective Compounds are described herein.
  • HIF ⁇ hydroxylation has been shown to result in therapeutic benefits. See, e.g., Freeman et ah, 2003; Welsh et ah, 2003; Siddiq et ah, 2005.
  • the purpose of the modulation ⁇ e.g., interaction, disruption, interference) of the hydroxylation of HIF ⁇ may be to, for example, induce stasis and/or modulate cellular functions such as angiogenesis, erythropoiesis, energy metabolism, inflammation, matrix metabolism, vasomotor function, and apoptotic/proliferative responses and pathophysiological responses to ischemia/hypoxia, all of which are mediated by HIF ⁇ as discussed herein.
  • the method of stabilizing the ⁇ -subunit of HIF comprises administering to a subject a compound that inhibits hydroxylation of HIF ⁇ .
  • the method comprises administering to a subject a compound that inhibits 2-oxoglutarate dioxygenase enzyme activity.
  • the 2-oxoglutarate dioxygenase enzyme is selected from the group consisting of EGLNl, EGLN2, EGLN3, EGLN9 (also called Egl-9), procollagen prolyl 4-hydroxylase, procollagen prolyl 3 -hydroxylase, procollagen lysyl hydroxylase, PHD4, FIH-I, and any subunit or fragment thereof, respectively.
  • the methods comprise inhibiting HIF prolyl hydroxylase enzyme activity.
  • the HIF prolyl hydroxylase enzyme is selected from the group consisting of EGLNl, EGLN2, EGLN3, EGLN9, and any subunit or fragment thereof, respectively.
  • the compound stabilizes HIF ⁇ by specifically inhibiting hydroxylation of at least one amino acid residue in HIF ⁇ .
  • the amino acid residue is selected from the group consisting of pro line and asparagine.
  • the agent inhibits hydroxylation of the HIF- l ⁇ P564 residue or a homologous proline in another HIF ⁇ isoform. In another specific embodiment, the agent inhibits hydroxylation of the HIF- l ⁇ P402 residue or a homologous proline in another HIF ⁇ isoform. In yet another embodiment, the compound may additionally inhibit hydroxylation of one or more HIF ⁇ asparagine residues. In one specific embodiment, the agent inhibits hydroxylation of the HIF- l ⁇ N 803 residue or a homologous asparagine residue in another HIF ⁇ isoform.
  • Methods for treating, preventing, or pretreating a HIF-associated condition in a subject comprising inhibiting 2-oxoglutarate dioxygenase enzyme activity, are also provided, and include methods in which the HIF-associated condition is one associated with ischemia, hypoxia, or stasis, or any other condition described herein.
  • the present invention provides a method for treating, preventing, or pretreating a HIF-associated condition, the method comprising administering to the subject a compound that inhibits 2-oxoglutarate dioxygenase enzyme activity.
  • the present invention provides a method of treating, preventing, or pretreating a HIF-associated condition in a subject, the method comprising inhibiting HIF prolyl hydroxylase enzyme activity.
  • HIF-associated conditions include those associated with hypoxia, ischemia, or with stasis, etc., as described herein.
  • the method comprises administering to the subject a compound that inhibits HIF prolyl hydroxylase activity.
  • the method further comprises administering a second compound.
  • the second compound inhibits 2-oxoglutarate dioxygenase enzyme activity, or the compound and the second compound inhibit the activities of different 2-oxoglutarate dioxygenase enzymes, or the second compound is selected from the group consisting of an ACE inhibitor (ACEI), angiotensin-II receptor blocker (ARB), diuretic, digoxin, statin, or carnitine, etc.
  • ACEI ACE inhibitor
  • ARB angiotensin-II receptor blocker
  • HIF-associated conditions include disorders such as pulmonary disorders, e.g., pulmonary embolism, etc., cardiac disorders, e.g., myocardial infarction, congestive heart failure, etc.
  • Acute ischemic events can include those associated with surgery, organ transplantation, infarction ⁇ e.g., cerebral, intestinal, myocardial, pulmonary, etc.), trauma, insult, or injury, etc.
  • Chronic events associated with ischemia can include hypertension, diabetes, occlusive arterial disease, chronic venous insufficiency, Raynaud's disease, cirrhosis, congestive heart failure, systemic sclerosis, etc.
  • the HIF-associated condition induces stasis.
  • treatment of an HIF-associated condition with an Effective Compound, as described herein induces stasis.
  • the present invention provides methods, compositions, articles of manufacture, and apparatuses to induce stasis in cells, tissues and organs located within or derived from an organism, as well as in the organism itself. Such methods, compositions, articles of manufacture, and apparatuses can be employed to protect biological matter, as well as to prevent, treat, or diagnose diseases and conditions in the organism.
  • such methods may directly induce stasis themselves, or they may act indirectly by not inducing stasis themselves, but by enhancing the ability of biological matter to enter stasis in response to an injury or disease condition, e.g., by reducing the time or level of injury or disease required to achieve stasis.
  • an injury or disease condition e.g., by reducing the time or level of injury or disease required to achieve stasis.
  • Such a condition may be referred to as pre- stasis. Details of such applications and other uses are described below.
  • the invention provides methods of pretreating or preconditioning wherein HIF ⁇ is stabilized prior to the occurrence of an event associated with a HIF- associated condition, e.g., ischemia, etc., or the development of a HIF-associated condition.
  • Ischemias can be induced by acute events. Such events can include, for example, surgery, e.g., angioplasty, organ transplantation, etc., and related procedures such as administration of anesthesia, etc.
  • the methods of pretreating or preconditioning are applied in situations where a subject has a disorder predictive of the development of a HIF-associated condition, e.g., transient ischemic attack or angina pectoris, indicative of stroke and myocardial infarction, respectively, in order to prevent the development of or reduce the degree of development of the HIF-associated condition.
  • a compound that stabilizes HIF ⁇ is administered to a subject in order to increase preconditioning factors for ischemia, for example, EPO, etc.
  • methods and/or compounds of the present invention are used to induce stasis or pre-stasis in biological matter, e.g., cells, tissues, organs, and/or organisms, after an injury (e.g., a traumatic injury) or after the onset or progression of a disease, in order to protect the biological matter from damage associated with the injury or disease prior to, during, or following treatment of the injury or disease.
  • methods of the present invention are used to induce or promote stasis or pre-stasis in biological matter prior to subjection to an injurious event (e.g., an elective surgery) or prior to the onset or progression of a disease, in order to protect the biological matter from damage associated with adverse conditions such as injury or disease.
  • Pre-treatment may include methods wherein biological matter is provided with an Effective Compound both before and during, and before, during and after biological matter is subjected to adverse conditions (e.g., an injury or onset or the progression of a disease), and methods wherein biological matter is provided with an Effective Compound only before biological matter is subjected to adverse conditions.
  • adverse conditions e.g., an injury or onset or the progression of a disease
  • the methods of the invention are used to prevent tissue damage caused by HIF-associated disorders including, but not limited to, ischemic and hypoxic disorders.
  • treatment is predicated on predisposing conditions, e.g., hypertension, diabetes, occlusive arterial disease, chronic venous insufficiency, Raynaud's disease, cirrhosis, congestive heart failure, and/or systemic sclerosis.
  • the methods of the invention can be used as a pretreatment to decrease or prevent the tissue damage caused by HIF-associated disorders including, but not limited to, ischemic and hypoxic disorders.
  • the need for pretreatment is based on a patient's history of recurring episodes of an ischemic condition, e.g., myocardial infarction or transient ischemic attacks, or has symptoms of impending ischemia, e.g., angina pectoris, etc.
  • the need for pretreatment is based on physical parameters implicating possible or likely ischemia or hypoxia, such as is the case with, e.g., individuals placed under general anesthesia or temporarily working at high altitudes.
  • the methods may be used in the context of organ transplants to pretreat organ donors and to maintain organs removed from the body prior to implantation in a recipient.
  • the present invention provides a method for increasing expression of angiogenic factors in a subject, the method comprising stabilizing HIF ⁇ .
  • the present invention provides a method of increasing expression of glycolytic factors in a subject, the method comprising stabilizing HIF ⁇ .
  • the invention provides a method of increasing expression of factors associated with oxidative stress in a subject, the method comprising stabilizing HIF ⁇ .
  • a method of treating a subject having a disorder associated with ischemic reperfusion injury, the method comprising stabilizing HIF ⁇ is also contemplated.
  • the present invention provides a method of identifying a compound that stabilizes HIF ⁇ , the method comprising: (a) administering a compound of interest to a subject or to a sample from a subject; (b) measuring the HIF ⁇ level in the subject or in the sample; and (c) comparing the HIF ⁇ level in the subject or in the sample to a standard level, wherein an increase in the HIF ⁇ level in the subject or the sample is indicative of a compound that stabilizes HIF ⁇ .
  • the invention provides compounds that stabilize HIF ⁇ and methods of using the compounds to prevent, pretreat, or treat HIF-associated conditions such as those described herein.
  • a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof, alone or in combination with a pharmaceutically acceptable excipient is administered to a subject having a HIF-associated condition.
  • the compound is administered immediately following the diagnosis of an acute ischemic disorder.
  • the compound is administered to a subject during the course of a chronic ischemic condition.
  • the ischemia is due to a transient or acute trauma, insult, or injury such as, e.g., a spinal cord injury.
  • the compound is administered to a patient in need following diagnosis of a pulmonary disorder such as COPD and the like.
  • the compound can be administered based on predisposing conditions, e.g., chronic conditions, or as a pretreatment to decrease or prevent tissue damage caused by HIF-associated disorders.
  • the compound is administered to a subject who has a history of recurring episodes of an ischemic condition, e.g. , myocardial infarction or transient ischemic attacks, or has symptoms of impending ischemia, e.g., angina pectoris.
  • the compound is administered based on physical parameters implicating possible ischemia or hypoxia, such as is the case with, e.g., individuals placed under general anesthesia or temporarily working at high altitudes.
  • the compounds may be used in the context of organ transplants to pretreat organ donors and to maintain organs removed from the body prior to implantation in a recipient.
  • a compound of the present invention induces stasis, as described herein. Stasis has been found to be modulated by HIF and also affects HIF prolyl hydroxylase activity (e.g., Egl-9). Therefore, the present invention provides methods, compositions, articles of manufacture, and apparatuses to induce stasis via modulation of HIF in cells, tissues and organs located within or derived from an organism, as well as in the organism itself. Such methods, compositions, articles of manufacture, and apparatuses can be employed to protect biological matter, as well as to prevent, treat, or diagnose diseases and conditions in the organism.
  • such methods may directly induce stasis themselves, or they may act indirectly by not inducing stasis themselves, but by enhancing the ability of biological matter to enter stasis in response to an injury or disease condition, e.g., by reducing the time or level of injury or disease required to achieve stasis.
  • an injury or disease condition e.g., by reducing the time or level of injury or disease required to achieve stasis.
  • Such a condition may be referred to as pre-stasis. Details of such applications and other uses are described below.
  • the invention is based, in part, on studies with compounds that were determined to have a protective function, and thus, serve as protective agents via modulation of HIF. Moreover, the overall results of studies involving different compounds indicate that compounds with an available electron donor center are particularly effective in inducing stasis or pre-stasis. In addition, these compounds induce reversible stasis, meaning they are not so toxic to the particular biologic matter that the matter dies or decomposes. It is further contemplated that the present invention can be used to enhance survivability of and/or to prevent or reduce damage to biological matter, which may be subject to or under adverse conditions.
  • methods of the present invention are used to induce stasis or pre-stasis in biological matter, e.g., cells, tissues, organs, and/or organisms, after an injury ⁇ e.g., a traumatic injury) or after the onset or progression of a disease, in order to protect the biological matter from damage associated with the injury or disease prior to, during, or following treatment of the injury or disease.
  • biological matter e.g., cells, tissues, organs, and/or organisms
  • an injury e.g., a traumatic injury
  • onset or progression of a disease e.g., a traumatic injury
  • methods of the present invention are used to induce or promote stasis or pre-stasis in biological matter prior to subjection to an injurious event ⁇ e.g., an elective surgery) or prior to the onset or progression of a disease, in order to protect the biological matter from damage associated with adverse conditions such as injury or disease.
  • Such methods are generally referred to as "pre-treatment" with an Effective Compound.
  • Pre-treatment includes methods wherein biological matter is provided with an Effective Compound both before and during, and before, during and after biological matter is subjected to adverse conditions ⁇ e.g., an injury or onset or the progression of a disease), and methods wherein biological matter is provided with an Effective Compound only before biological matter is subjected to adverse conditions.
  • stasis may be induced by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with an Effective Compound that induces stasis directly itself or, alternatively, by treating biological matter with
  • Effective Compound that does not itself induce stasis, but instead, promotes or enhances the ability of or decreases the time required for the biological matter to achieve stasis in response to another stimuli, such as, but not limited to, an injury, a disease, hypoxia, excessive bleeding, or treatment with another Effective Compound.
  • treatment with an Effective Compound induces
  • pre-stasis which refers to a hypometabolic state through which biological matter must transition to reach stasis.
  • Pre-stasis is characterized by a reduction in metabolism within the biological material of a magnitude that is less than that defined as stasis. Further, pre-stasis can be considered an HIF-associated condition.
  • the biological matter In order to achieve stasis using an Effective Compound, the biological matter necessarily must transition through a graded hypometabolic state in which oxygen consumption and CO 2 production are reduced less than two-fold in the biological matter. Such a continuum, in which metabolism or cellular respiration is reduced by an Effective Compound
  • stasis comprises a two-fold reduction (i.e., a reduction to 50% or less) in either CO 2 production or O 2 consumption
  • direct measurement of these parameters in the biological matter using methods known to those in the art in which a reduction of less than two-fold is detected is indicative of pre-stasis.
  • certain measurements of carbon dioxide and oxygen levels in the blood as well as other markers of metabolic rate familiar to those skilled in the art including, but not limited to, blood p ⁇ 2 , VO 2 , pCO 2 , pH, and lactate levels, may be used in the instant invention to monitor the onset or progression of pre-stasis.
  • pre-stasis may be associated with an at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% reduction in CO 2 evolution, which refers to the amount of CO 2 released from the lungs.
  • pre-stasis is characterized by a redution in one or more indicators of metabolic activity that is less than or equal to 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 49% as compared to normal physiological conditions.
  • pre-stasis is characterized by its ability to enhance or promote entry into stasis in response to another stimuli (wherein the another stimuli may include prolonged treatment with the same active agent), or its ability to enhance survival of or protect biological matter from damage resulting from an injury, the onset or progression of the disease, or bleeding, particularly bleeding that can lead to irreversible tissue damage, hemorrhagic shock, or lethality.
  • methods of the present invention explicitly exemplified herein may refer to inducing "stasis,” it is understood that these methods may be readily adapted to induce “pre-stasis,” and that such methods of inducing pre-stasis are contemplated by the present invention.
  • the same Effective Compounds used to induce stasis may also be used to induce pre-stasis, by providing them to biological matter at, for example, a lower dosage and/or for a shorter time than used to induce stasis.
  • the present invention involves exposing biological matter to an amount of an agent, so as to achieve stasis of the biological matter via modulation of HIF.
  • the present invention concerns methods for inducing stasis in in vivo biological matter comprising: a) identifying an organism in which stasis is desired; and, b) exposing the organism to an effective amount of an Effective Compound, such as an oxygen antagonist, to induce stasis in the in vivo biological matter.
  • “Inducing stasis” in biological matter means that the matter is alive but is characterized by one or more of the following: at least a two-fold reduction in the rate or amount of carbon dioxide production by the biological matter; at least a two-fold ⁇ i.e., 50%) reduction in the rate or amount of oxygen consumption by the biological matter; and at least a 10% decrease in movement or motility (applies only to cells or tissue that move, such as sperm cells or a heart or a limb, or when stasis is induced in the entire organism) (collectively referred to as "cellular respiration indicators").
  • embodiments of the invention may be discussed in terms of a reduction in the rate of oxygen consumption by the biological matter as about, at least about, or at most about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any range derivable therein.
  • any assay to measure oxygen consumption may be employed, and a typical assay will involve utilizing a closed environment and measuring the difference between the oxygen put into the environment and oxygen that is left in the environment after a period of time. It is further contemplated that carbon dioxide production can be measured to determine the amount of oxygen consumption by biological matter. Thus, there may be decreases in carbon dioxide production, which would correspond to the decreases in oxygen consumption discussed above.
  • stasis or pre-stasis is temporary and/or reversible, meaning that the biological matter no longer exhibits the characteristics of stasis at some later point in time.
  • a compound that is not does not qualify as an oxygen antagonist is administered. It is contemplated that methods discussed with respect to oxygen antagonists may be applied with respect to any compound that is an oxygen antagonist, protective metabolic agent, compound with the structure of Formula I, Ia- Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X, XI, any other compound discussed herein, or a salt or precursor thereof.
  • induction of stasis is desired in which case the compound may be referred to as an "effective stasis compound.” It is contemplated that in some embodiments of the invention, a method is achieved by inducing stasis. For example, therapeutic methods may involve inducing stasis, in which case the Effective Compound is an effective stasis compound. It is specifically contemplated that in embodiments in which Effective Compounds are discussed, the invention includes, and may be limited to, oxygen antagonists.
  • biological matter is treated with an Effective Compound that does not induce stasis by itself (at least not at the level and/or duration of time provided), but rather induces biological matter to enter a pre-stasis state that has therapeutic benefits and that enhances the ability of the biological matter to achieve stasis in response to another stimuli, such as, e.g., an injury, disease state, or treatment with another Effective Compound or the same Effective Compound if used for a longer duration or greater dosage.
  • Another stimuli such as, e.g., an injury, disease state, or treatment with another Effective Compound or the same Effective Compound if used for a longer duration or greater dosage.
  • biological matter refers to any living biological material (such as mammalian biological material, in some embodiments) including cells, tissues, organs, and/or organisms, and any combination thereof.
  • stasis may be induced in a part of an organism (such as in cells, in tissue, and/or in one or more organs), whether that part remains within the organism or is removed from the organism, or the whole organism will be placed in a state of stasis.
  • m vivo biological matter refers to biological matter that is in vivo, i.e., still within or attached to an organism.
  • biological matter will be understood as synonymous with the term “biological material.”
  • one or more cells, tissues, or organs is separate from an organism.
  • isolated can be used to describe such biological matter. It is contemplated that stasis may be induced in isolated biological matter.
  • An organism or other biological matter in need of stasis is an organism or biological matter in which stasis of all or part of the organism may yield direct or indirect physiological benefits.
  • a patient at risk for hemorrhagic shock may be considered in need of stasis, or a patient who will undergo coronary artery bypass surgery may benefit from protecting the heart from ischemia/reperfusion injury.
  • Other applications are discussed throughout the application.
  • an organism or other biological matter is identified or determined to be in need of stasis based on one or more tests, screens, or evaluations that indicate a condition or disease, or the risk of a condition or disease that can be prevented or treated by undergoing stasis.
  • the taking of a patient medical or family medical history may yield information that an organism or other biological matter is in need of stasis.
  • one application of the present invention would be to reduce the overall energy demands of a biological material by inducing stasis.
  • an organism or other biological matter may be in need of an Effective Compound to enhance survivability.
  • a patient may need treatment for an injury or disease or any other application discussed herein. They may be determined to be in need of enhanced survivability or treatment based on methods discussed in the previous paragraph, such as by taking a patient medical or family medical history.
  • an effective amount is one that induces stasis in the tissue or organ as determined by the collective amount of cellular respiration of the tissue or organ. Accordingly, for example, if the level of oxygen consumption by a heart (collectively with respect to cells of the heart) is decreased at least about 2-fold (i.e., 50%) after exposure to a particular amount of a certain Effective Compound, such as an oxygen antagonist or an effective stasis compound, it will be understood that that was an effective amount to induce stasis in the heart. Similarly, an effective amount of an agent that induces stasis in an organism is one that is evaluated with respect to the collective or aggregate level of a particular parameter of stasis.
  • an effective amount when inducing stasis in an organism, an effective amount is one that induces stasis generally of the whole organism, unless a particular part of the organism was targeted.
  • an effective amount may be an amount sufficient to induce stasis by itself, or it may be an amount sufficient to induce stasis in combination with another agent or stimuli, e.g., another Effective Compound, an injury, or a disease state.
  • Another agent or stimuli e.g., another Effective Compound, an injury, or a disease state.
  • the concept of an effective amount of a particular compound is related, in some embodiments, to how much utilizable oxygen there is available to the biological matter.
  • stasis can be induced when there is about 100,000 ppm or less of oxygen in the absence of any oxygen antagonist (room air has about 210,000 ppm oxygen).
  • the oxygen antagonist serves to alter how much oxygen is effectively available. At concentration of 10 ppm of oxygen, suspended animation is induced. Thus, while the actual concentration of oxygen that biological matter is exposed to may be higher, even much higher, than 10 ppm, stasis can be induced because of the competitive effect of an oxygen antagonist with oxygen for binding to essential oxygen metabolizing proteins in the biological matter. In other words, an effective amount of an oxygen antagonist reduces the effective oxygen concentration to a point where the oxygen that is present cannot be used. This will happen when the amount of an oxygen antagonist reduces the effective oxygen concentration below the K m of oxygen binding to essential oxygen metabolizing proteins (i.e., comparable to 10 ppm of oxygen).
  • an oxygen antagonist reduces the effective concentration of oxygen by about or at least about 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 15-, 20-, 25-, 30-, 35-, 40-, 45-, 50-, 60-, 70-, 80-, 90-, 100-, 150-, 200-, 250-, 300-, 350-, 400-, 450-, 500-, 600-, 700-, 800-, 900-, 1000-, HOO-, 1200-, 1300-, 1400-, 1500-, 1600-, 1700-, 1800-, 1900-, 2000-, 2100-, 2200-, 2300-, 2400-, 2500-, 2600-, 2700-, 2800-, 2900-, 3000- , 3100-, 3200-, 3300, 3400-, 3500-, 3600-, 3700-, 3800-, 3900-, 4000-, 4100-, 4200-, 4300-, 4400-, 4500-, 5000-,
  • embodiments of the invention may be discussed in terms of a reduction in effective oxygen concentration as about, at least about, or at most about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any range derivable therein.
  • stasis can be measured indirectly by a drop in core body temperature of an organism. It is contemplated that a reduction in core body temperature of about, at least about, or at most about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 0 F or more, or any range derivable therein may be observed in methods of the invention.
  • hypothermia can be induced, such as moderate hypothermia (at least 1O 0 F reduction) or severe hypothermia (at least 2O 0 F reduction).
  • the effective amount can be expressed as a concentration with or without a qualification on length of time of exposure.
  • an oxygen antagonist or other Effective Compound for about, at least about, or at most about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 seconds, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5,
  • the amount of time may be indefinite, depending on the reason or purpose for administering the oxygen antagonist or other Effective Compound.
  • biological matter may continue to be exposed to an oxygen antagonist or other Effective Compound, or, in other embodiments of the invention, the biological matter may no longer be exposed to the oxygen antagonist or other Effective Compound. This latter step can be achieved either by removing or effectively removing the oxygen antagonist or other Effective Compound from the presence of the biological matter in which stasis was desired, or the biological matter may be removed from an environment containing the oxygen antagonist or other Effective Compound.
  • matter may be exposed to or provided with any Effective Compound continuously (a period of time without a break in exposure), intermittently (exposure on multiple occasions), or on a periodic basis (exposure on multiple occasions on a regular basis).
  • the dosages of the Effective Compound on these different bases may the same or they may vary.
  • an Effective Compound is provided periodically by providing or exposing biological matter to an Effective Compound 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8,
  • biological matter is exposed to or provided with an Effective Compound for a sustained period of time, where "sustained” means a period of time of at least about 2 hours.
  • biological matter may be exposed to or provided with an Effective Compound for a sustained period of time, where "sustained” means a period of time of at least about 2 hours.
  • biological matter may be exposed to or provided with an Effective Compound for a sustained period of time, where "sustained” means a period of time of at least about 2 hours.
  • biological matter may be exposed to or provided with an Effective composition compound for a sustained period of time, where "sustained” means a period of time of at least about 2 hours.
  • biological matter is provided with an Effective Compound on a continuously sustained basis.
  • biological matter may be exposed to or provided with an Effective Compound for about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more hours (or any range derivable therein) for 2, 3, 4, 5, 6, 7 days, and/or 1, 2, 3, 4, 5 weeks, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9,
  • biological matter may be exposed to or provided with an Effective Compound at least before and during; before, during, and after; during and after; or solely after a particular injury, trauma, or treatment (for instance, surgery), adverse condition or other relevant event or situation. This exposure may or may not be sustained.
  • the dosages of the Effective Compound on these different bases may the same or they may vary.
  • an Effective Compound may be provided on a continously sustained basis at level that is considered "low,” meaning a level that is less than the amount that causes metabolic flexibility such as seen with drop in CBT, heart rate, or CO 2 or O 2 consumption or production.
  • biological matter is exposed or provided an Effective Compound
  • Compound such as a metabolic agent, in an amount that exceeds what was previously understood to be the maximum tolerated dose before adverse physiological ramifications such as apnea ("period of time during which breathing is markedly reduced such that the subject takes 10% or fewer number of breaths"), lack of observable skeletal muscle movement, dystonia, and/or hyperactivity.
  • apnea peripheral of time during which breathing is markedly reduced such that the subject takes 10% or fewer number of breaths
  • Such an amount may be particularly relevant to increasing survivability in some embodiments of the invention, for instance, to increase the chances of surviving adverse conditions, such as those that would induce death from hemorrhagic shock.
  • a physiological state can be induced by Effective Compounds of the present invention which enhances survivability in an organism in need of survivability enhancement and comprises a set of observable physiological changes in response to an effective dose of an Effective Compound, said changes may comprise one, more or all of hyperpnea, apnea and the concomitant or subsequent loss of neuromuscular tone or voluntary control of movement with continued heartbeat.
  • hyperpnea refers to rapid, shallow breathing.
  • Apnea refers to a cessation of breathing or the reduction as described above.
  • the subject becomes apneic, which is marked by a cessation in breathing and then an apnic breath after a short period of time. In rats, this occurs after approximately 20 seconds.
  • a subject induced into apnea may exhibit 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10% the number of breaths subsequent to exposure to an Effective Compound.
  • the subject may have an occasional breath, which may be considered an apnic breath, thereafter.
  • apnea continues until the subject is no longer exposed to the Effective Compound.
  • an effective amount may be expressed as LD 5O , which refers to the "median lethal dose,” which means the dose that is administered that kills half the population of animals (causes 50% mortality).
  • an effective amount may be independent of the weight of the biological matter ("weight independent").
  • weight independent In rodents and humans, for example, the LD 50 Of H 2 S gas is approximately 700 ppm before adverse physiological effects occur.
  • increasing survivability refers generally to living longer, which is an embodiment of the invention.
  • the present invention also concerns methods for inducing apnea in an organism comprising administering to the organism an effective amount of an Effective Compound.
  • the organism also does not exhibit any skeletal muscle movement as a result of the Effective Compound. It is specifically contemplated that the organism may be mammal, including a human.
  • an effective amount exceeds what is considered a lethal concentration.
  • the concentration may be a lethal amount though the exposure time may be about, at least about, or at most about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 seconds, 1, 2, 3, 4, 5 minutes or more (or any range derivable therein or any other time period specified in this disclosure).
  • a mammal is exposed to at least about 600 ppm of an Effective Compound in a gaseous form, such as H 2 S.
  • an Effective Compound in a gaseous form such as H 2 S.
  • there is a step of identifying an animal in need of treatment in other embodiments, there is a step of observing apnea in the organism.
  • methods involving obtaining a blood sample from the organism and/or evaluating the color of the organism's blood It has been observed that exposure to H 2 S changes the color of blood from a mammal; it goes from bright red to a darker, red wine color and then to brick red.
  • Evaluating the color may be done visually without any instruments or machines, while in other embodiments, an instrument may be used, such as a spectrophotometer.
  • a blood sample may be obtained from an organism and other types of analysis may be done on it.
  • a blood sample may not be needed and instead, blood may be evaluated without the sample.
  • a modified pulse-oximeter that shines IR or visible light through the finger may be employed to monitor color changes in the blood.
  • biological matter is exposed to an effective amount of an Effective Compound that does not lead to stasis or pre-stasis. In some embodiments, there may be no evidence of a reduction in oxygen consumption or carbon dioxide production while the Effective Compound is present.
  • an organism may be exposed to the Effective Compound while sleeping.
  • the exposure may be regular, such as daily (meaning exposure at least once a day).
  • an Effective Compound is provided to a subject by nebulizer. This may be applied with any embodiment of the invention.
  • the nebulizer is used for the treatment of hemorrhagic shock.
  • the Effective Compound is provided as a single dose to the subject.
  • a single dose or multiple doses is one that would induce apnea in a subject.
  • a subject is given at least about 1,000, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 11,000, 12,000 or more ppm H 2 S gas.
  • the exposure time may be any of the times discussed herein, including about or about at most 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1 minutes or less (or any range derivable therein).
  • the metabolic rate of biologic matter may change.
  • the RQ ratio (CO 2 production/O 2 consumption) of the biological matter changes after exposure to an Effective Compound. This may occur after an initial exposure or repeated exposure or after an acute exposure.
  • the RQ ratio decreases after exposure.
  • the decrease may be a decrease of about, at least about or at most about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80% or more, or any range derivable therein.
  • the decrease may be a result of O 2 consumption increasing of CO 2 production decreasing in relation to O 2 consumption.
  • methods involve measuring an RQ ratio in a subject. This may occur before and/or after exposure to the Effective Compound.
  • stasis is induced, and a further step in methods of the invention is to maintain the relevant biological matter in a state of stasis. This can be accomplished by continuing to expose the biological matter to an oxygen antagonist or other Effective Compound and/or exposing the biological matter to a nonphysiological temperature or another oxygen antagonist or other Effective Compound. Alternatively, the biological matter may be placed in a preservation agent or solution, or be exposed to normoxic or hypoxic conditions.
  • biological matter may be maintained in stasis for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or range derivable therein.
  • a change in pressure or to effect a cryoprotectant or cryopreservation environment e.g., one containing glycerol
  • a cryoprotectant or cryopreservation environment e.g., one containing glycerol
  • stasis with respect to a whole animal and “stasis” with respect to cells or tissues may require different lengths of time in stasis.
  • a time of stasis of up to 12, 18, or 24 hours is generally contemplated.
  • non-human animal subjects e.g. non-human animals shipped or stored for commercial purposes, stasis for a period of 2 or 4 days, 2 or 4 weeks, or longer is contemplated.
  • exposure is used according to its ordinary meaning to indicate that biological matter is subjected to an oxygen antagonist or other Effective Compound. This can be achieved in some embodiments by contacting biological matter with an oxygen antagonist or Effective Compound. In other embodiments, this is achieved by contacting the biological matter with an Effective Compound, which may or may not be an oxygen antagonist. In the case of in vivo cells, tissues, or organs, “expose” may further mean “to lay open” these materials so that it can be contacted with an oxygen antagonist or other Effective Compound. This can be done, for example, surgically.
  • Exposing biological matter to an oxygen antagonist or other Effective Compound can be by incubation in or with (includes immersion) the antagonist, perfusion or infusion with the antagonist, injection of biological matter with an oxygen antagonist or other Effective Compound, or applying an oxygen antagonist or other Effective Compound to the biological matter.
  • stasis of the entire organism is desirable, inhalation or ingestion of the oxygen antagonist or other Effective Compound, or any other route of pharmaceutical administration is contemplated for use with oxygen antagonists or other Effective Compound.
  • the term "provide” is used according to its ordinary and plain meaning to mean “to supply.” It is contemplated that a compound may be provided to biological matter in one form and be converted by chemical reaction to its form as an Effective Compound.
  • an effective amount is characterized as a sublethal dose of the oxygen antagonist or other Effective Compound.
  • a “sublethal dose” means a single administration of the oxygen antagonist or Effective Compound that is less than half of the amount of the oxygen antagonist or Effective Compound that would cause at least a majority of cells in a biological matter to die within 24 hours of the administration.
  • a "sublethal dose” means a single administration of the oxygen antagonist or Effective Compound that is less than half of the amount of the oxygen antagonist or Effective Compound that would cause the organism to die within 24 hours of the administration. In other embodiments, an effective amount is characterized as a near-lethal dose of the oxygen antagonist or Effective Compound.
  • a “near lethal dose” means a single administration of the oxygen antagonist or Effective Compound that is within 25% of the amount of the inhibitor that would cause at least a majority of cell(s) to die within 24 hours of the administration.
  • a "near lethal dose” means a single administration of the oxygen antagonist or Effective Compound that is within 25% of the amount of the inhibitor that would cause the organism to die within 24 hours of the administration.
  • a sublethal dose is administered by administering a predetermined amount of the oxygen antagonist or Effective Compound to the biological material. It is specifically contemplated that this may be implemented with respect to any Effective Compound.
  • an effective amount is characterized as a supralethal dose of the oxygen antagonist or other Effective Compound.
  • a "supra-lethal dose” means a single administration of an Effective Compound that is at least 1.5 times (1.5x) the amount of the Effective Compound that would cause at least a majority of cells in a biological matter to die within 24 hours of the administration. If stasis of the entire organism is desired, then a "supra-lethal dose” means a single administration of the Effective Compound that is at least 1.5 times the amount of the Effective Compound that would cause the organism to die within 24 hours of the administration.
  • the supra- lethal dose can be about, at least about, or at most about 1.5x, 2x, 3x, 4x, 5x, 1Ox, 2Ox, 30x, 4Ox, 5Ox, 6Ox, 7Ox, 80x, 9Ox, 10Ox, 15Ox, 20Ox, 25Ox, 300x, 40Ox, 50Ox, 60Ox, 70Ox, 800x, 90Ox, 100Ox, 110Ox, 120Ox, 1300x, 140Ox, 150Ox, 160Ox, 170Ox, 1800x, 190Ox, 200Ox, 3000x, 400Ox, 500Ox, 600Ox, 700Ox, 8000x, 900Ox, 10,000x or more, or any range derivable therein, the amount of the Effective Compound that would cause at least a majority of cells in a biological matter (or the entire organism) to die within 24 hours of the administration.
  • the amount of the Effective Compound that is provided to biological matter can be about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220,
  • the amount may be expressed as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280,
  • an effective amount is administered by monitoring, alone or in combination, the amount of oxygen antagonist or other Effective Compound administered, monitoring the duration of administration of the oxygen antagonist or other Effective Compound, monitoring a physiological response (e.g., pulse, respiration, pain response, movement or motility, metabolic parameters such as cellular energy production or redox state, etc.) of the biological material to the administration of the oxygen antagonist or other Effective Compound and reducing, interrupting or ceasing administration of the compound(s) when a predetermined floor or ceiling for a change in that response is measured, etc.
  • a physiological response e.g., pulse, respiration, pain response, movement or motility, metabolic parameters such as cellular energy production or redox state, etc.
  • these steps can be employed additionally in any method of the invention.
  • Tissue in a state of stasis or that has undergone stasis can be used in a number of applications. They can be used, for example, for transfusion or transplantation (therapeutic applications, including organ transplants); for research purposes; for screening assays to identify, characterize, or manufacture other compounds that induce stasis; for testing a sample from which the tissue was obtained (diagnostic applications); for preserving or preventing damage to the tissue that will be placed back into the organism from which they were derived (preventative applications); and for preserving or preventing damage to them during transport or storage. Details of such applications and other uses are described below.
  • isolated tissue means that the tissue is not located within an organism. In some embodiments, the tissue is all or part of an organ.
  • tissue and "organ” are used according to their ordinary and plain meanings. Though tissue is composed of cells, it will be understood that the term “tissue” refers to an aggregate of similar cells forming a definite kind of structural material. Moreover, an organ is a particular type of tissue.
  • the present invention concerns methods for inducing stasis in isolated tissue comprising: a) identifying the tissue in which stasis is desired; and, b) exposing the tissue to an effective amount of an oxygen antagonist or other Effective Compound to induce stasis.
  • compositions, methods, and articles of manufacture of the invention can be used on biological matter that will be transferred back into the donor organism from which it was derived (autologous) or a different recipient (heterologous) subject.
  • biological matter is obtained directly from a donor organism.
  • the biological matter is placed in culture prior to exposure to an oxygen antagonist or other Effective Compound.
  • the biological matter is obtained from a donor organism administered extracorporeal membrane oxygenation prior to retrieval of the biological matter, which is a technique implemented to aid in the preservation of biological matter.
  • methods include administering or implanting the biological matter in which stasis was induced to a live recipient organism.
  • an organ or tissue to be retrieved and then transplanted is exposed to the oxygen antagonist or other Effective Compound while still in the donor subject. It is contemplated that in some cases, the vasculature of the donor is used to expose the organ or tissue to the oxygen antagonist or other Effective Compound. This can be done if the heart is still pumping or a pump, catheter, or syringe can be used to administer the oxygen antagonist or other Effective Compound into the vasculature for delivery to the organ or tissue
  • Methods of the invention also concern inducing stasis in isolated tissue comprising incubating the tissue with an oxygen antagonist or other effective stasis compound that creates hypoxic conditions for an effective amount of time for the tissue to enter stasis.
  • Cells in a state of stasis or that have undergone stasis can be used in a number of applications. They can be used, for example, for transfusion or transplantation
  • therapeutic applications for research purposes; for screening assays to identify, characterize, or manufacture other compounds that induce stasis; for testing a sample from which the cells were obtained (diagnostic applications); for preserving or preventing damage to the cells that will be placed back into the organism from which they were derived (preventative applications); and for preserving or preventing damage to cells during transport or storage. Details of such applications and other uses are described below.
  • the present invention concerns methods for inducing stasis in one or more cells separate from an organism comprising: a) identifying the cell(s) in which stasis is desired; and, b) exposing the cell(s) to an effective amount of an oxygen antagonist or other effective stasis compound to induce stasis.
  • the cell may be any oxygen-utilizing cell.
  • the cell may be eukaryotic or prokaryotic.
  • the cell is eukaryotic. More particularly, in some embodiments, the cell is a mammalian cell. Mammalian cells contemplated for use with the invention include, but are not limited to those that are from a: human, monkey, mouse, rat, rabbit, hamster, goat, pig, dog, cat, ferret, cow, sheep, and horse.
  • cells of the invention may be diploid but in some cases, the cells are haploid (sex cells). Additionally, cells may be polyploid, aneuploid, or anucleate.
  • the cell can be from a particular tissue or organ, such as one from the group consisting of: heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood, small intestine, large intestine, brain, spinal cord, smooth muscle, skeletal muscle, ovary, testis, uterus, and umbilical cord.
  • the cell can also be characterized as one of the following cell types: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, barrier function cell, contractile cell, absorptive cell, mucosal cell, limbus cell (from cornea), stem cell (totipotent, pluripotent or multipotent), unfertilized or fertilized oocyte, or sperm.
  • cell types platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, barrier function cell, contractile cell, absorptive cell, mucosal cell, limbus cell (from cornea), stem cell (totipotent, pl
  • the present invention also provides methods, compositions, and apparati for enhancing survivability of and/or reducing damage to biological matter under adverse conditions by reducing metabolic demand, oxygen requirements, temperature, or any combination thereof in the biological matter of interest.
  • survivability of biological matter is enhanced by providing it with an effective amount of a protective metabolic agent.
  • the agent enhances survivability by preventing or reducing damage to the biological matter, preventing all or part of the matter from dying or senescing, and/or extending the lifespan of all or part of the biological matter, relative to biological matter not exposed to the agent.
  • the agent prolongs survival of tissue and/or an organism that would otherwise not survive without the agent.
  • a "protective metabolic agent” is a substance or compound capable of reversibly altering the metabolism of biological matter that is exposed to or contacted with it and that promotes or enhances the survivability of the biological matter.
  • the protective metabolic agent induces stasis in the treated biological matter; while, in other embodiments, the protective metabolic agent does not directly itself induce stasis in the treated biological matter.
  • Protective metabolic agents, and other Effective Compounds may enhance survivability and/or reduce damage to biological matter without inducing stasis in the biological matter per se, but rather by reducing cellular respiration and corresponding metabolic activity to a degree that is less than about a fifty percent decrease in oxygen consumption or carbon dioxide production. Additionally, such compounds may cause the biological matter to more quickly, easily, or effectively enter into or achieve stasis in response to an injury or disease state, e.g., by inducing the biological matter to achieve a state of pre-stasis.
  • Survivability includes survivability when the matter is under adverse conditions —that is, conditions under which there can be adverse and nonreversible damage or injury to biological matter.
  • Adverse conditions can include, but are not limited to, when oxygen concentrations are reduced in the environment (hypoxia or anoxia, such as at high altitudes or under water); when the biological matter is incapable of receiving that oxygen (such as during ischemia), which can be caused by i) reduced blood flow to organs (e.g., heart, brain, and/or kidneys) as a result of blood vessel occlusion (e.g., myocardial infarction, and/or stroke), ii) extracorporeal blood shunting as occurs during heart/lung bypass surgery (e.g., "pumphead syndrome" in which heart or brain tissue is damaged as a result of cardiopulmonary bypass), or iii) as a result of blood loss due to trauma (e.g., hemorrhagic shock or surgery); hypothermia, where the biological material is subjected
  • a protective metabolic agent is an oxygen antagonist in certain embodiments of the invention. It is also contemplated that in certain other embodiments, an oxygen antagonist is not a protective metabolic agent.
  • one or more compounds may be used to increase or enhance survivability of biological matter; reversibly inhibit the metabolism and/or activity of biological matter; reduce the oxygen requirement of biological matter; reduce or prevent damage to biological matter under adverse conditions; prevent or reduce damage or injury to biological matter; prevent aging or senescence of biological matter; and, provide a therapeutic benefit as described throughout the application with respect to oxygen antagonists. It is contemplated that embodiments relating to inducing stasis are applicable to these other embodiments as well. Therefore, any embodiment discussed with respect to stasis may be implemented with respect to these other embodiments.
  • An Effective Compound used for inducing stasis or any of these other embodiments may lead or provide their desired effect(s), in some embodiments, only when they are in the context of the biological matter (i.e., have no lasting effect) and/or they may provide for these effect(s) for more than 24 hours after the biological matter is no longer exposed to it. Moreover, this can also be the case when a combination of Effective Compounds is used.
  • biological matter is exposed to an amount of an oxygen antagonist or other Effective Compound that reduces the rate or amount of carbon dioxide production by the biological matter at least 2-fold, but also by about, at least about, or at most about 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 15-, 20-, 25-, 30-, 35-, 40-,
  • embodiments of the invention may be discussed in terms of a reduction in the rate or amount of carbon dioxide production by the biological matter as about, at least about, or at most about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any range derivable therein.
  • biological matter is exposed to an amount of an oxygen antagonist or other Effective Compound that reduces the rate or amount of oxygen consumption by the biological matter at least 2- fold, but also by about, at least about, or at most about 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 15- , 20-, 25-, 30-, 35-, 40-, 45-, 50-, 100-, 200-, 300-, 400-, 500-fold of more, or any range derivable therein.
  • an oxygen antagonist or other Effective Compound that reduces the rate or amount of oxygen consumption by the biological matter at least 2- fold, but also by about, at least about, or at most about 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 15- , 20-, 25-, 30-, 35-, 40-, 45-, 50-, 100-, 200-, 300-, 400-, 500-fold of more, or any range derivable therein.
  • embodiments of the invention may be discussed in terms of a reduction in the rate or amount of oxygen consumption by the biological matter as about, at least about, or at most about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any range derivable therein.
  • biological matter is exposed to an amount of an oxygen antagonist or other Effective Compound that decreases movement or motility by at least 10%, but also by about, at least about, or at most about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, or 100%, or any range derivable therein.
  • these characteristics and parameters are in the context of whichever biological matter is induced into a state of stasis. Thus, if stasis is induced in an organism's heart, these parameters would be evaluated for the heart, and not the whole organism.
  • a reduction in oxygen consumption on the order of roughly 8- fold is a kind of stasis referred to as "hibernation.”
  • a reduction in oxygen consumption on the order of around 1000- fold can be considered “suspended animation.”
  • embodiments of the invention concerning stasis can be achieved at the level of hibernation or suspended animation, if appropriate.
  • a "-fold reduction" is relative to the reduced amount; for example, if a non-hibernating animal consumes 800 units of oxygen, the hibernating animal consumes 100 units of oxygen.
  • methods are provided for reducing cellular respiration, which may or may not be as high as that needed to reach stasis.
  • a reduction in oxygen consumption by about, at least about, or at most about 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% is provided in methods of the invention. This can also be expressed and assessed in terms of any cellular respiration indicator. It is contemplated that biological matter may be exposed to one or more oxygen antagonists or other Effective Compounds more than one time.
  • biological matter may be exposed to one or more Effective Compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times, meaning when a biological matter is exposed multiple times that there are periods of respite (with respect to exposure to the Effective Compound) in between.
  • an Effective Compound may be administered before, during, after, or any combination thereof, in relation to the onset or progression of an injurious insult or disease condition.
  • pre- treatment of biological matter to an Effective Compound is sufficient to enhance survivability and/or reduce damage from an injurious or disease insult.
  • Pre-treatment is defined as exposure of the biological matter to the Effective Compound before the onset or detection of the injurious or disease insult.
  • Pre-treatment can be followed by termination of exposure at or near the onset of the insult or continued exposure after the onset of insult.
  • methods including pre-exposure to an Effective composition including pre-exposure to an Effective
  • Compound i.e., pre-treatment are used to treat conditions in which an injurious or disease insult is 1) scheduled or elected in advance, or 2) predicted in advance to likely occur.
  • Examples meeting condition 1 include, but are not limited to, major surgery where blood loss may occur spontaneously or as a result of a procedure, cardiopulmonary bypass in which oxygenation of the blood may be compromised or in which vascular delivery of blood may be reduced (as in the setting of coronary artery bypass graft (CABG) surgery), or in the treatment of organ donors prior to removal of donor organs for transport and transplantation into a recipient in need of an organ transplant.
  • CABG coronary artery bypass graft
  • Examples meeting condition 2 include, but are not limited to, medical conditions in which a risk of injury or disease progression is inherent (e.g., in the context of unstable angina, following angioplasty, bleeding aneurysms, hemorrhagic strokes, following major trauma or blood loss), or in which the risk can be diagnosed using a medical diagnostic test.
  • Exposure to the Effective Compound may enhance survivability or reduce damage when exposure occurs after the onset or detection of the injurious or disease insult to achieve a therapeutic effect. Exposure to the Effective Compound may be brief or extended.
  • the exposure duration may be only for as long as needed to reach an indicator of stasis activity or pre-stasis (e.g., blood pCO 2 , p ⁇ 2 , pH, lactate, or sulfhemoglobin levels, or body temperature), or it may be longer.
  • exposure occurs following traumatic injury (including iatrogenic and/or non-iatro genie injuries) to an organism and is used to induce stasis or pre-stasis in the entire organism or injured tissue therein, so as to prevent or minimize damage, e.g., ischemic and reperfusion injury prior to, during, and/or following treatment.
  • the present invention includes a method of protecting a mammal from suffering cellular damage from a surgery, comprising providing to the mammal an amount of hydrogen sulfide or other Effective Compound sufficient to induce the mammal to enter pre-stasis prior to the surgery.
  • the surgery may be elective, planned, or emergency surgery, such as, e.g., cardiopulmonary surgery.
  • the hydrogen sulfide may be administered by any means available in the art, including, e.g., intravenously or by inhalation.
  • the present invention includes a method of protecting a mammal from suffering cellular damage from a disease or adverse medical condition, comprising providing to the mammal an amount of hydrogen sulfide or other Effective Compound sufficient to induce the mammal to enter pre-stasis or stasis prior to the onset or progression of the disease or adverse medical condition.
  • This embodiment may be used in the context of a variety of different diseases and adverse medical conditions, including, e.g., unstable angina, post-angioplasty, aneurism, hemorrhagic stroke or shock, trauma, and blood loss.
  • the invention concerns methods of preventing an organism, such as a mammal, from bleeding to death or suffering irreversible tissue damage as a result of bleeding by providing to the mammal an amount of hydrogen sulfide or other Effective Compound sufficient to prevent the animal from bleeding to death.
  • the organism may go into hemorrhagic shock but not die from excessive bleeding.
  • bleeding and “hemorrhaging” are used interchangeably to refer to any discharge of blood from a blood vessel. It includes, but is not limited to, internal and external bleeding, bleeding from an injury (which may be from an internal source, or from an external physical source such as from a gunshot, stabbing, physical trauma, etc.).
  • additional embodiments of the invention concern prevention of death or irreversible tissue damage from blood loss or other lack of oxygenation to cells or tissue, such as from lack of an adequate blood supply.
  • This may be the result of, for example, actual blood loss, or it may be from conditions or diseases that prevent cells or tissue from being perfused (e.g., reperfusion injury), that cause blockage of blood to cells or tissue, that reduce blood pressure locally or overall in an organism, that reduce the amount of oxygen is carried in the blood, or that reduces the number of oxygen carrying cells in the blood.
  • Conditions and diseases that may be involved include, but are not limited to, blood clots and embolisms, cysts, growths, tumors, anemia (including sickle cell anemia), hemophilia, other blood clotting diseases (e.g., von Willebrand, ITP), and atherosclerosis.
  • Such conditions and diseases also include those that create essentially hypoxic or anoxic conditions for cells or tissue in an organism because of an injury, disease, or condition.
  • a sublethal collective dose or a nonlethal collective dose is administered to the biological matter.
  • a "sublethal collective dose” means an amount of multiple administrations of the Effective Compound that collectively is less than half of the amount of the Effective Compound that would cause at least a majority of cell(s) to die within 24 hours of one of the administrations.
  • an effective amount is characterized as a near-lethal dose of the oxygen antagonist or other Effective Compound.
  • a “near lethal collective dose” means an amount of multiple administrations of the oxygen antagonist or other Effective Compound that is within 25% of the amount of the Effective Compound that would cause at least a majority of cell(s) to die within 24 hours of the one of the administrations.
  • a “supra-lethal collective dose” means an amount of multiple administrations of the Effective Compound that is at least 1.5 times the amount of the Effective Compound that would cause at least a majority of cell(s) (or the entire organism) to die within 24 hours of the one of the administrations. It is contemplated that multiple doses can be administered so as to induce stasis in the whole organism.
  • the definition for "sub-lethal collective dose,” “near-lethal collective dose” and “supra-lethal collective dose” can be extrapolated based on the individual doses discussed earlier for stasis in whole organisms.
  • biological matter may be exposed to or contacted with more than one oxygen antagonist or other Effective Compound.
  • Biological matter may be exposed to at least one Effective Compound, including 2, 3, 4, 5, 6, 7, 8, 9, 10 or more oxygen antagonists or other Effective Compound, or any range derivable therein.
  • the term "effective amount" refers to the collective amount of Effective Compounds.
  • the biological matter may be exposed to a first Effective Compound and then exposed to a second Effective Compound.
  • biological matter may be exposed to more than one Effective Compound at the same time or in an overlapping manner.
  • more than one Effective Compounds may be comprised or mixed together, such as in a single composition to which biological matter is exposed. Therefore, it is contemplated that, in some embodiments, a combination of Effective Compounds is employed in compositions, methods, and articles of manufacture of the invention.
  • Biological matter may be provided with or exposed to an Effective Compound through inhalation, injection, catheterization, immersion, lavage, perfusion, topical application, absorption, adsorption, or oral administration.
  • biological matter may be provided with or exposed to an Effective Compound by administration to the biological matter intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostaticaly, intrapleurally, intratracheally, intranasally, intrathecally, intravitreally, intravaginally, intrarectally, topically, intratumorally, intramuscularly, intraperitoneally, intraocularly, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion, via a catheter, or via a lavage.
  • Methods and apparatuses of the invention involve a protective agent that in some embodiments is an oxygen antagonist.
  • the oxygen antagonist is a reducing agent.
  • the oxygen antagonist can be characterized as a chalcogenide compound. It will be understood that Effective Compounds may also be protective agents. Moreover, any chalcogenide compound can be considered an Effective Compound so long as it achieves a goal of the invention, regardless of whether it is an oxygen antagonist.
  • the chalcogenide compound comprises sulfur, while in others, it comprises selenium, tellurium, or polonium.
  • a chalcogenide compound contains one or more exposed sulfide groups. It is contemplated that this chalcogenide compounds contains 1, 2, 3, 4, 5, 6 or more exposed sulfide groups, or any range derivable therein. In particular embodiments, such a sulf ⁇ de-containing compound is CS 2 (carbon disulfide).
  • biological matter is provided with a precursor compound that becomes the active version of any compound of the present invention by exposure to biological matter, such as by chemical or enzymatic means.
  • the compound may be provided to the biological matter as a salt of the compound in the form of a free radical, or a negatively charged, positively charged or multiply charged species.
  • Some compounds qualify as both a Formula I and a Formula IV compound and in such cases, the use of the phrase "Formula I or Formula IV" is not intended to connote the exclusion of such compounds.
  • a compound identified by the structure of Formula I or Formula IV may also, in certain embodiments, be characterized as an oxygen antagonist, protective metabolic agent, or a precursor, prodrug, or salt thereof. It is further contemplated that the compound need not be characterized as such or qualify as such to be a compound used in the invention, so long as it achieves a particular method of the invention. In some other embodiments, the compound may be considered a chalcogenide compound. It is specifically contemplated that any compound identified by the structure of Formula I or Formula
  • any compound set forth in this disclosure may be used instead of or in addition to an oxygen antagonist in methods, compositions, and apparatuses of the invention; similarly, any embodiments discussed with respect to any of structure having Formula I or Formula IV or otherwise set forth in this disclosure may be may be used instead of or in addition to an oxygen antagonist. Moreover, any compound set forth in this disclosure may be combined with any oxygen antagonist or any other
  • more than one compound with the structure of Formula I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X, XI, or any other compound of the present invention is provided.
  • multiple different compounds with a structure from the same formula e.g., Formula I or Formula IV
  • the amount of carbon dioxide to which the biological matter may be exposed is about, at least about, or at most about , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30% or more, or any range derivable therein.
  • the amount is expressed in terms of ppm, such as about, at least about, or at most about 350, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 6000, 7000, 8000, 9000, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000, 20000, 21000, 22000, 23000, 24000, 25000, 26000, 27000, 28000, 29000, 30000, 31000, 32000, 33000, 34000, 35000, 36000, 37000, 35000,
  • the Effective Compound is sodium sulfide, sodium thiomethoxide, cysteamine, sodium thiocyanate, cysteamine-S-phosphate sodium salt, or tetrahydrothiopyran-4-ol.
  • the Effective Compound is dimethylsulfoxide, thioacetic acid, selenourea, 2-(3-aminopropyl)-aminoethanethiol-dihydrogen-phosphate-ester, 2- mercapto-ethanol, thioglycolicether, sodium selenide, sodium methane sulfmate, thiourea, or dimethylsulf ⁇ de.
  • any subset of Effective Compounds identified by name or structure may be used in methods, compositions and articles of manufacture. It is also specifically contemplated that any subset of these compounds may be disclaimed as not constituting embodiments of the invention.
  • the present invention also concerns pharmaceutical compositions comprising a therapeutically effective amount of one or more Effective Compounds. It is understood that such pharmaceutical compositions are formulated in pharmaceutically acceptable compositions.
  • the composition may include a pharmaceutically acceptable diluent.
  • the pharmaceutical composition contains an effective dose of an active to provide when administered to a patient a C max or a steady state plasma concentration of the Effective Compound to produce a therapeutically effective benefit.
  • the C max or steady state plasma concentration to be achieved is about, at least about, or at most about 0.01, 0.1, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200
  • the desired C max or steady state plasma concentration is about between 10 ⁇ M to about 10 mM, or between about 100 ⁇ M to about 1 mM, or between about 200 ⁇ M to about 800 ⁇ M.
  • Appropriate measures may be taken to consider and evaluate levels of the compound already in the blood, such as sulfur.
  • the pharmaceutical composition provides an effective dose of H 2 S to provide when administered to a patient a C max or a steady state plasma concentration of between 10 ⁇ M to 10 mM, between about 100 ⁇ M to about 1 mM, or between about 200 ⁇ M to about 800 ⁇ M.
  • the dosing of the salt is based on administering approximately the same sulfur equivalents as the dosing of the H 2 S. Appropriate measures will be taken to consider and evaluate levels of sulfur already in the blood.
  • the composition comprises a gaseous form of one or more of the Effective Compounds specified herein.
  • the composition comprises a salt of one or more of these compounds.
  • a pharmaceutical composition comprises a gaseous form of Formula I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X or XI, or a salt of Formula I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X or XI.
  • a gaseous form or salt of H 2 S is specifically contemplated in some aspects of the invention.
  • the amount of gas to which biological matter is provided is about, at least about, or at most about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840
  • the effective amount of gas(es) may be expressed as about, at least about, or at most about 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
  • the amount of gas to which biological matter is provided is about, at least about, or at most about 5, 10, 15,
  • the pharmaceutical composition is a liquid.
  • the composition may be a liquid with the relevant compound(s) dissolved or bubbled into the composition.
  • the pharmaceutical composition is a medical gas.
  • gases are those gases that are drugs within the meaning of ⁇ 201(g)(l) of the Federal Food, Drug and Cosmetic Act (“the Act") (21 U.S.C. ⁇ 321(g) and pursuant to ⁇ 503(b)(l)(A) of the Act (21 U.S.C. ⁇ 353(b)(l)(A) are required to be dispensed by prescription.
  • the Act 21 U.S.C. ⁇ 353(b)(l)(A)
  • a medical gas includes at least one Effective Compound.
  • the present invention further comprises apparatuses and articles of manufacture comprising packaging material and, contained within the packaging material, an effective stasis compound, wherein the packaging material comprises a label that indicates that it can be used for inducing stasis in in vivo biological matter.
  • the apparatus or article of manufacture further includes a pharmaceutically acceptable diluent.
  • the apparatus or article of manufacture has a buffering agent.
  • the Effective Compound is provided in a first sealed container and the pharmaceutically acceptable diluent is provided in a second sealed container.
  • the device or article further has instructions for mixing the Effective Compound and the diluent.
  • the Effective Compound can be reconstituted for achieving any method of the invention, such as for inducing stasis in in vivo biological matter. It is contemplated that any label would specify the result to be achieved and the use of the compound for patients in need of that result.
  • the present invention also concerns an article of manufacture comprising packed together: an Effective Compound, instructions for use of the effective stasis compound, comprising: (a) identifying in vivo tissue in need of stasis treatment; and (b) administering an effective amount of the Effective Compound to the in vivo biological matter.
  • an article of manufacture comprising a medical gas including an Effective Compound and a label comprising details or use and administration for inducing stasis in a biological matter or any other method of the invention.
  • kits for the delivery of an Effective Compound to a tissue site in need of stasis treatment, or any other treatment of the claimed invention comprising: a drape adapted for forming a sealed envelope against a tissue site; a container comprising an Effective Compound; and an inlet in the drape, wherein the container comprising the Effective Compound is in communication with the inlet.
  • the kit includes an outlet in the drape wherein the outlet is in communication with a negative pressure source.
  • the drape comprises elastomeric material and/or has a pressure sensitive adhesive covering the periphery of the drape. The outlet may be placed in fluid communication with the negative pressure source, which may or may not be a vacuum pump.
  • the kit includes a canister, which may or may not be removable, in fluid communication between the outlet and the negative pressure source. It is contemplated that the container includes an Effective Compound that is in gaseous communication with the inlet. In certain embodiments, the container includes an Effective Compound that is a gas or a liquid gas.
  • the kit may also include a vaporizer in communication between the container comprising an oxygen antagonist and the inlet. In addition, it may have a return outlet in communication with the container comprising the Effective Compound.
  • the Effective Compound in the kits is carbon monoxide, carbon dioxide, H 2 Se, and/or H 2 S.
  • the tissue site for which the kit or method is applied is wounded.
  • any compound discussed herein as an oxygen antagonist can be provided in prodrug form to the biological matter, meaning that the biological matter or other substance(s) in the environment of the biological matter alters the prodrug into its active form, that is, into an oxygen antagonist. It is contemplated that the term "precursor" covers compounds that are considered "prodrugs.”
  • the oxygen antagonist or other Effective Compound may be or may be provided as a gas, semi-solid liquid (such as a gel or paste), liquid, or solid. It is contemplated that biological matter may be exposed to more than one such Effective Compound
  • Effective Compound may be formulated for a particular mode of administration, as is discussed herein.
  • the Effective Compound is in pharmaceutical acceptable formulation for intravenous delivery.
  • the Effective Compound is a gas.
  • the gaseous Effective Compound includes carbon monoxide, carbon dioxide, nitrogen, sulfur, selenium, tellurium, or polonium, or a mixture thereof.
  • the Effective Compound may, in certain embodiments, be a chalcogenide compound as a gas.
  • the Effective Compound is in a gas mixture comprising more than one gas.
  • the other gas(es) is a non-toxic and/or a non-reactive gas in some embodiments.
  • the other gas is a noble gas (helium, neon, argon, krypton, xenon, radon, or ununoctium), nitrogen, nitrous oxide, hydrogen, or a mixture thereof.
  • the non-reactive gas may simply be a mixture that constitutes "room air," which is a mixture of nitrogen, oxygen, argon and carbon dioxide, as well as trace amounts of other atoms/molecules such as neon, helium, methane, krypton, and hydrogen. The precise amounts of each varies, though a typical sample might contain about 78% nitrogen, 21% oxygen, 0.9% argon, and 0.04% carbon dioxide.
  • room air is a mixture containing about 75 to about 81% nitrogen, about 18 to about 24% oxygen, about 0.7 to about 1.1% argon, and about 0.02% to about 0.06% carbon dioxide.
  • a gaseous Effective Compound may be first diluted with a non-toxic and/or non-reactive gas prior to administration or exposure to biological matter. Additionally or alternatively, any gaseous Effective Compound may be mixed with room air prior to administration or exposure to biological matter or the compound may be administered or exposed to the biological matter in room air. In some instances, the gas mixture also contains oxygen.
  • Compound gas is mixed with oxygen to form an oxygen gas (O 2 ) mixture in other embodiments of the invention.
  • an oxygen gas mixture in which the amount of oxygen in the oxygen gas mixture is less than the total amount of all other gas or gases in the mixture.
  • the Effective Compound gas is carbon monoxide and the amount of carbon monoxide is about the same or exceeds any amount of oxygen in the oxygen gas mixture.
  • carbon monoxide is employed with blood-free biological matter.
  • blood-free biological matter refers to cells and organs whose oxygenation is not dependent, or no longer dependent, on the vasculature, such as an organ for transplant.
  • the atmosphere may be 100% CO, but as will be evident to one skilled in the art, the amount of CO may be balanced with gases other than oxygen providing that the amount of usable oxygen is reduced to a level that prevents cellular respiration.
  • the ratio of carbon monoxide-to- oxygen may be 85:15 or greater, 199:1 or greater or 399:1 or greater.
  • the ratio is about, at least about, or at most about 1 :1, 2:1, 2.5:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1. 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, 100:1, 110:1, 120:1, 130:1, 140:1, 150:1, 160:1, 170:1, 180:1, 190:1, 200:1, 210: 1, 220:1, 230:1, 240:1, 250:1, 260:1, 270:1, 280:1, 290:1, 300:1, 310:1, 320:1, 330: 1, 340:1, 350:1, 360:1, 370:1, 380:1, 390:1, 400:1, 410:1, 420:1, 430:1, 440:1, 450: 1, 460:1, 470:1, 480:1, 490:1, 500:1 or more, or any range
  • the above numbers pertain to the ratio of carbon monoxide to a mixture of oxygen and one or more other gases.
  • the other gas is a nonreactive gas such as nitrogen (N 2 ).
  • N 2 nitrogen
  • the above numbers apply to ratios of carbon monoxide to a combination of oxygen and nitrogen (O 2 /N 2 ) that can be used in methods and apparatuses of the invention. Accordingly, it will be understood that other gases may or may not be present.
  • the CO:oxygen ratio is balanced with one or more other gases (non-carbon monoxide and non-oxygen gases). In particular embodiments, the CO:oxygen ratio is balanced with nitrogen.
  • the amount of CO is a ratio of CO compared to room air, as is described by the numbers above.
  • the amount of carbon monoxide is relative to the amount of oxygen, while in others, it is an absolute amount.
  • the amount of oxygen is in terms of "parts per million (ppm)" which is a measure of the parts in volume of oxygen in a million parts of air at standard temperature and pressure of 20 0 C and one atmosphere pressure and the balance of the gas volume is made up with carbon monoxide.
  • ppm parts per million
  • the amount of carbon monoxide to oxygen is related in terms of parts per million of oxygen balanced with carbon monoxide.
  • the atmosphere to which the biological material is exposed or incubated may be at least 0, 50, 100, 200, 300, 400, 500, 1000, or 2000 parts per million (ppm) of oxygen balanced with carbon monoxide and in some cases carbon monoxide mixed with a non-toxic and/or non-reactive gas
  • the term "environment" refers to the immediate environment of the biological matter, that is, the environment with which it is in direct contact.
  • the biological material must be directly exposed to carbon monoxide, and it is insufficient that a sealed tank of carbon monoxide be in the same room as the biological matter and be considered to be incubated an "environment" according to the invention.
  • the atmosphere may be expressed in terms of kPa.
  • the environment in which a biological material is incubated or exposed to is about, at least about, or at most about 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.
  • the atmosphere is about, at least about, or at most about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 101, 101.3 kPa CO, or any range derivable therein.
  • the partial pressure is about or at least about 85, 90, 95, 101, 101.3 kPa CO, or any range derivable therein.
  • the amount of time the sample is incubated or exposed to carbon monoxide can also vary in embodiments of the invention.
  • the sample is incubated or exposed to carbon monoxide for about, for at least about, or for at most about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 or more minutes and/or, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, and/or 1, 2, 3, 4, 5, 6,7, 8, 9, 10 or more days.
  • the invention concerns compositions and articles of manufacture that contain one or more Effective Compounds.
  • a composition has one or more of these Effective Compounds as a gas that is bubbled in it so that the composition provides the compound to the biological matter when it is exposed to the composition.
  • Such compounds may be gels, liquids, or other semisolid material.
  • a solution has an oxygen antagonist as a gas bubbled through it. It is contemplated that the amount bubbled in the gas will provide the appropriate amount of the compound to biological material exposed to the solution. In certain embodiments, the amount of gas bubbled into the solution is about, at least about, or at most about 0.5, 1.0, 1.5, 2.0.
  • Biological matter is exposed to the gas in a closed container in some embodiments of the invention.
  • the closed container can maintain a particular environment or modulate the environment as is desired.
  • the environment refers to the amount of oxygen antagonist that the biological matter is exposed and/or the temperature, gas composition, or pressure of the environment.
  • the biological matter is placed under a vacuum before, during, or after exposure to an oxygen antagonist or other Effective Compound. Moreover, in other cases, the biological matter is exposed to a normoxic environment after being exposed to an oxygen antagonist or other Effective Compound.
  • the present invention includes methods for inducing stasis or protecting biological matter from injury or disease that include providing an Effective Compound to the biological matter in combination with providing another stasis-inducing Effective Compound or environmental condition to the biological matter. Such combination treatment may occur in any order, e.g., simultaneously or sequentially.
  • an Effective Compound is provided to biological matter, and the biological matter is subsequently placed under hypoxic conditions, such as 5% O 2 , or sequentially exposed to increasingly hypoxic conditions, such as 5% O 2 followed by 4% O 2 , 3%
  • the environment containing the biological matter cycles at least once to a different amount or concentration of the oxygen antagonist or other Effective Compound, wherein the difference in amount or concentration is by at least one percentage difference.
  • the environment may cycle back and forth between one or more amounts or concentrations of the oxygen antagonist or other Effective Compound, or it may gradually increase or decrease the amount or concentrations of an that compound.
  • the different amount or concentration is between about 0 and 99.9% of the amount or concentration of the oxygen antagonist or other Effective Compound to which the biological matter was initially exposed. It is contemplated that the difference in amount and/or concentration is about, at least about, or at most about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2,
  • Methods of the invention can also include a step of subjecting biological matter to a controlled temperature environment.
  • the biological matter is exposed to a temperature that is a "nonphysiological temperature environment,” which refers to a temperature in which the biological matter cannot live in for more than 96 hours.
  • the controlled temperature environment can have a temperature of about, at least about, or at most about -210, -200, -190, -180, -170, - 160, -150, -140, -130, -120, -110, -100, -90, -80, -70, -60, -50, -40, -30, -20, -10, -5, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82,
  • Biological matter may also be exposed to an oxygen antagonist or other Effective Compound at room temperature, which means a temperature between about 2O 0 C and about 25 0 C. Furthermore, it is contemplated the biological matter achieves a core temperature of any amount or range of amounts discussed.
  • the biological matter can be subjected to a nonphysiological temperature environment or a controlled temperature environment before, during or after exposure to the oxygen antagonist(s) or other Effective Compound(s). Furthermore, in some embodiments, the biological matter is subjected to a nonphysiological temperature environment or a controlled temperature environment for a period of time between about one minute and about one year.
  • the amount of time may be about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or range derivable therein.
  • the temperature may be altered or cycled during the process in which temperature is controlled.
  • the temperature of the biological matter may first be reduced before it is placed in the environment that has the oxygen antagonist or other Effective Compound, while in others, the biological matter may be cooled by placing it in the environment with the
  • Methods of the invention can also include a step of subjecting biological matter to a controlled pressure environment.
  • the biological matter is exposed to pressure that is lower than the pressure under which the organism is typically under.
  • the biological matter is subjected to a "nonphysiological pressure environment," which refers to a pressure under which the biological matter cannot live under for more than 96 hours.
  • the controlled pressure environment can have a pressure of about, at least about, or at most 10 ⁇ 14 , 10 ⁇ 13 , 10 ⁇ 12 , 10 "11 , 10 "10 , 10 “9 , l ⁇ 10 "7 , l ⁇ 10 ⁇ 5 , l ⁇ 10 ⁇ 3 , l ⁇ 10 "1 , 0.2, 0.3, 0.4 or 0.5 atm or more, or any range derivable therein. It is contemplated that the biological matter can be subjected to a nonphysiological pressure environment or a controlled pressure environment before, during or after exposure to the Effective Compound(s).
  • the biological matter is subjected to a nonphysiological pressure environment or a controlled pressure environment for a period of time between about one minute and about one year.
  • the amount of time may be about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1,
  • the pressure may be altered or cycled during the process in which pressure is controlled.
  • the pressure to which the biological matter is exposed may first be reduced before it is placed in the environment that has the Effective Compound, while in others, the biological matter placed under pressure after exposure to an Effective Compound.
  • the pressure may be reduced gradually, such that the pressure of the environment starts at one pressure but then reaches another pressure within 10, 20, 30, 40, 50, 60 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, and/or 1, 2, 3, 4, 5, 6, 7 days or more, and any combination or range derivable therein.
  • methods include modulating environmental oxygen levels or removing the biological material from an environment having oxygen.
  • exposing biological material to an environment in which oxygen is diminished or absent may mimic exposure of the biological material to an oxygen antagonist.
  • biological matter is exposed to or provided with an Effective Compound under conditions in which the environment of the biological matter is hypoxic or anoxic, as described in further detail below. This may be intentional or nonintentional.
  • biological matter is intentionally placed in an environment that is anoxic or hypoxic or in an environment that is made anoxic or hypoxic.
  • the biological matter is under such conditions as a result of an unintended situation, for example, if the biological matter is under ischemic or potentially ischemic conditions. Therefore, it is contemplated in some cases that the hypoxic or anoxic conditions would damage the matter in the absence of the Effective Compound.
  • toxicity can be controlled for by altering the level, amount, duration, or frequency of an Effective Compound and/or environmental change to which the biological matter is exposed.
  • the alteration is a reduction, while in certain other embodiments, the alteration is an increase. It is contemplated that the skilled artisan is aware of a number of ways of evaluating toxicity effects in biological matter.
  • compositions, methods, and articles of manufacture of the invention can be used on biological matter that will be transferred back into the donor organism from which it was derived (autologous) or a different recipient (heterologous) subject.
  • biological matter is obtained directly from a donor organism.
  • the biological matter is placed in culture prior to exposure to an oxygen antagonist or other Effective Compound.
  • the biological matter is obtained from a donor organism administered extracorporeal membrane oxygenation prior to retrieval of the biological matter, which is a technique implemented to aid in the preservation of biological matter.
  • methods include administering or implanting the biological matter in which stasis was induced to a live recipient organism.
  • Methods of the invention also concern inducing stasis in in vivo biological matter comprising incubating the biological matter with an oxygen antagonist or other Effective Compound that creates hypoxic conditions for an effective amount of time for the biological matter to enter stasis.
  • oxygen demand is reduced about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95
  • aspects of the invention concern methods for preserving in vivo biological matter comprising exposing the in vivo biological matter to an effective amount of an oxygen antagonist or other Effective Compound to preserve the biological matter in vivo.
  • the present invention also concerns a method of delaying or reducing the effects of trauma on or in an organism comprising exposing biological matter at risk for trauma to an effective amount of an oxygen antagonist or other Effective Compound.
  • there are methods for treating or preventing hemorrhagic shock in a patient comprising exposing the patient to an effective amount of an oxygen antagonist or other Effective Compound.
  • methods prevents lethality in the patient as a result of the bleeding and/or hemorrhagic shock.
  • steps include exposing the patient to an effective amount of an oxygen antagonist or other Effective Compound.
  • the oxygen antagonist is specifically contemplated to be a chalcogenide compound such as H 2 S.
  • Methods for reducing heart rate in an organism are also included as part of the invention. Such methods involve contacting the biological sample or organism with an effective amount of an oxygen antagonist or other Effective Compound.
  • One embodiment of the invention relates to a method of inducing hibernation in a mammal comprising contacting the mammal with an effective amount of an oxygen antagonist or other Effective Compound.
  • a method of anesthetizing an organism comprising exposing biological matter in which anesthesia is desired to an effective amount of an oxygen antagonist or other Effective Compound. It is contemplated that the anesthesia may be similar to local or general anesthesia.
  • the present invention further includes methods of protecting a mammal from radiation therapy or chemotherapy comprising contacting the mammal with an effective amount of an oxygen antagonist or other Effective Compound prior to or during radiation therapy or chemotherapy.
  • an oxygen antagonist or other Effective Compound prior to or during radiation therapy or chemotherapy.
  • Compound may also be administered locally to the affected organ, tissue, and/or cells.
  • methods can be used for preventing or reducing hair loss in a chemotherapy patient. It is contemplated that such a patient may have already received chemotherapy or be a candidate for chemotherapy. In particular cases, it is contemplated that an Effective Compound is provided to the patient as a topical gel to be applied where the hair loss is anticipated or present.
  • a hyperproliferative disease e.g., cancer
  • contacting the mammal with an effective amount of an oxygen antagonist or other Effective Compound comprising contacting the mammal with an effective amount of an oxygen antagonist or other Effective Compound and subjecting the mammal to hyperthermia therapy.
  • methods of the invention may be applied to preserving organs for transplant, other aspects of the invention concern the recipient organism.
  • methods of inhibiting rejection of an organ transplant in a mammal comprising providing the mammal with an effective amount of an oxygen antagonist or other Effective Compound.
  • Temperature regulation can be achieved in organisms by employing oxygen antagonists or other Effective Compounds.
  • there is a method of treating a subject with hypothermia comprising (a) contacting the subject with an effective amount of an oxygen antagonist, and then (b) subjecting the subject to an environmental temperature above that of the subject.
  • the present invention includes a method of treating a subject with hyperthermia comprising (a) contacting the subject with an effective amount of an oxygen antagonist or other Effective Compound.
  • treatment of hyperthermia also includes (b) subjecting the subject to an environmental temperature that is at least about 2O 0 C below that of the subject.
  • exposing the subject to nonphysiological or a controlled temperature environment can be used in additional embodiments.
  • the invention concerns a method for inducing cardioplegia in a patient undergoing bypass surgery comprising administering to the patient an effective amount of an oxygen antagonist or other Effective Compound. It is contemplated that administration may be local to the heart so as to protect it.
  • aspects of the invention relate to a method for preventing hematologic shock in a patient comprising administering to the patient an effective amount of an oxygen antagonist or other Effective Compound.
  • methods for promoting wound healing in an organism comprising administering to the organism or wound an effective amount of an oxygen antagonist or other Effective Compound.
  • the present invention covers a method for preventing or treating neurodegeneration in a mammal comprising administering to the mammal an effective amount of an oxygen antagonist or other Effective Compound.
  • the present invention also covers reducing the oxygen requirement of biological matter, meaning that the amount of oxygen required by the biological matter to survive is reduced. This can be achieved by providing an effective amount of one or more Effective Compounds. It is generally known how much oxygen particular biological matter require to survive, which can also be dependent on time, pressure, and temperature.
  • the oxygen requirement of the biological matter is reduced by about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100%, or any range derivable therein, as compared to the requirement of the biological matter in the absence of the effective amount
  • Additional embodiments of the invention concern methods for preventing hair loss, such as from chemotherapy, by administering to a patient who has or will undergo chemotherapy an effective amount of at least one Effective Compound.
  • the biological matter may be exposed to an oxygen antagonist within about, within at least about, or within at most about 30 seconds, 1,
  • methods include an initial assessment of any damage, trauma, a wound, or degeneration.
  • a hematological disorder which means a disease, disorder or condition that affects any hematopoietic cells or tissue.
  • a hematological disorder which means a disease, disorder or condition that affects any hematopoietic cells or tissue. Examples include sickle cell disease and thalassemia.
  • CF cystic fibrosis
  • methods for enhancing survivability in a patient with cystic fibrosis (CF) by administering or providing an effective amount of an Effective Compound.
  • methods for treating cynanide poisoning in a subject comprising administering an effective amount of an Effective Compound.
  • the compound is H 2 S.
  • aspects of the invention concern methods for preserving one or more cells that are separate from an organism comprising contacting the cell(s) with an effective amount of an oxygen antagonist or other Effective Compound to preserve the one or more cells.
  • an oxygen antagonist or other Effective Compound to preserve the one or more cells.
  • shrimp embryos are specifically contemplated for use with the present invention.
  • oxygen reduction techniques can be embodied in a kit.
  • the kit currently sold under product number 261215 available from Becton Dickinson, makes use of select techniques described here.
  • That kit includes an anaerobic generator (e.g., a hydrogen gas generator), Palladium Catalysts, an anaerobic indicator, and a gas impermeable, sealable, "BioBag" into which the above components (together with platelets in a gas-permeable bag) are placed and sealed.
  • an anaerobic generator e.g., a hydrogen gas generator
  • Palladium Catalysts Palladium Catalysts
  • an anaerobic indicator an anaerobic indicator
  • a gas impermeable, sealable "BioBag” into which the above components (together with platelets in a gas-permeable bag) are placed and sealed.
  • rotenone is not the compound employed in this method, or possibly other methods of the invention. Moreover, it is also contemplated that in some embodiments, rotenone is excluded as an Effective Compound. Similarly, it is contemplated that nitric oxide may be excluded as an Effective Compound. In other embodiments of the invention, methods are provided for enhancing the ability of biological matter to enter stasis in response to an injury or disease by providing an effective amount of an Effective Compound, thereby protecting the biological matter from damage or injury, thereby enhancing survival of biological matter. Related embodiments include methods of preparing or priming biological matter for entry into stasis in response to an injury or disease by providing an effective amount of an Effective Compound.
  • Other related embodiments include method of inducing biological matter into pre-stasis, thereby protecting the biological matter from damage or injury.
  • treatment with an Effective Compound at a dosage or for a time less than required to induce stasis enables the biological matter to more readily or more completely achieve a beneficial state of stasis in response to an injury or disease, while in the absence of treatment with the Effective Compound, the biological matter would die or suffer damage or injury before it reached a protective level of stasis, e.g., a level sufficient to render the biological matter resistant to lethal hypoxia.
  • a protective level of stasis e.g., a level sufficient to render the biological matter resistant to lethal hypoxia.
  • hypoxia, ischemia, and blood loss all reduce the amount of oxygen available and supplied to oxygen utilizing biological matter, thereby reducing oxygen utilization in cells of the biological matter, reducing energy production derived from oxidative phosphorylation, and thereby decreasing thermogenesis, leading to hypothermia.
  • stasis may or may not have been achieved.
  • Treatment with an Effective Compound lowers the threshold (i.e., the severity or duration of the insult that is needed to achieve stasis) for induction of stasis, or it may add to or synergize with the injurious or disease stimuli to induce stasis in biological matter under injurious conditions that would not have resulted in stasis were it not for the Effective Compound treatment.
  • Such activity of Effective Compounds is determined by comparing the stasis-inducing effects (magnitude, kinetics) of injurious or disease stimuli alone with those in which the biological matter was pre-exposed, exposed concomitantly, exposed after, or any combination thereof, to the Effective Compound.
  • a method for inducing sleep in an organism comprising exposing the organism to an effective amount of an Effective Compound, wherein the effective amount is less than an amount that can induce stasis in the organism.
  • the term "sleep" is used according to its ordinary and plain meaning in a medical context. Sleep is distinguishable from other states of unconsciousness, which are also contemplated as states that can be achieved using methods of the invention.
  • the present invention also concerns methods for anesthetizing biological matter comprising exposing the matter to an effective amount of an Effective Compound, wherein the effective amount is less than an amount that can induce stasis in the organism.
  • an effective amount that is less than an amount that can induce stasis in an organism may be reduced with respect to duration and/or amount. That reduction may be a reduction in amount by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent, or any range derivable therein,
  • a reduction may be a reduction in duration (length of exposure time) by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7, days, 1, 2, 3, 4, 5 weeks, and/or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or any range derivable therein.
  • the reduction may be in terms of the overall effective amount provided to the biological matter, which may be a reduction of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 percent, or any range derivable therein, relative to the overall effective amount to induce stasis in an organism of that species and/
  • Methods of the invention can involve employing an apparatus or system that maintains the environment in which biological matter is placed or exposed to.
  • the invention includes an apparatus in which an oxygen antagonist or other Effective
  • the apparatus includes a container with a sample chamber for holding the biological matter, wherein the container is connected to a supply of gas comprising the oxygen antagonist(s).
  • the container may be a solid container or it may flexible, such as a bag.
  • the invention is an apparatus for preserving cell(s), the apparatus comprising: a container having a sample chamber with a volume of no greater than 775 liters; and a first gas supply in fluid communication with the sample chamber, the first gas supply including carbon monoxide.
  • the apparatus also includes a cooling unit that regulates the temperature inside the sample chamber and/or a gas regulator that regulates the amount of oxygen antagonist or other Effective Compound in the chamber or the amount of oxygen antagonist or other Effective Compound in a solution that is in the chamber. It is contemplated that there may be a gas supply for a second or additional gas or a second or additional gas supply for the oxygen antagonist or other Effective Compound.
  • the second gas supply may be connected with the sample chamber or it may be connected with the first gas supply.
  • the additional gas as discussed above, may be a non-toxic and/or non-reactive gas.
  • a gas regulator is part of the apparatus in some embodiments of the invention.
  • the gas regulator regulates the gas supplied to the sample chamber from the first gas supply. Alternatively, it regulates the gas supplied to the sample chamber or first gas supply from the second gas supply, or there may be a regulator for both the first and second gas supplies. It is further contemplated that any gas regulator can be programmed to control the amount of gas supplied to the sample chamber and/or to another gas supply. The regulation may or may not be for a specified period of time. There may be a gas regulator, which may or may not be programmable, for any gas supply directly or indirectly connected to the sample chamber. In some cases, the gas regulator is electronically programmable.
  • the pressure and/or the temperature inside the chamber can be regulated with either a pressure regulator or temperature regulator, respectively. As with the gas regulator, these regulators may be electronically programmable.
  • the apparatus of the invention may also have a cooling and/or heating unit to achieve the temperatures discussed above. The unit may or may not be electronically programmable.
  • the apparatus includes a wheeled cart on which the container rests or it may have one or more handles.
  • the invention includes an apparatus for cell(s), tissues, organs, and even whole organisms, in which the apparatus has: a container having a sample chamber; a first gas supply in fluid communication with the sample chamber, the first gas supply including the oxygen antagonist(s) or other Effective Compound(s); and an electronically-programmable gas regulator that regulates gas supplied to the sample chamber from the first gas supply.
  • the apparatus also has a structure configured to provide a vacuum within the sample chamber.
  • any oxygen antagonist or other Effective Compound described in this application is contemplated for use with apparatuses of the invention.
  • carbon monoxide can be administered using this apparatus.
  • a chalcogenide compound can be administered or a compound having the reducing agent structure.
  • an Effective Compound is administered using the apparatus.
  • the invention covers a device or its use.
  • the device is single dose delivery device.
  • the device is an inhaler or nebulizer.
  • other devices include, but are not limited to, an injection device such as a pen, a pump such as an infusion pump, or a patch. Moreover, it is contemplated that these devices may or may not be single dose delivery devices.
  • a candidate substance is screened for the ability to act as an oxygen antagonist or Effective Compound, specifically including a protective metabolic agent. This can be done using any assay described herein, such as by measuring carbon dioxide output. Any substance identified as having exhibiting characteristics of an oxygen antagonist or other Effective Compound can be further characterized or tested. Moreover, it is contemplated that such a substance can be administered to biological matter to induce stasis or manufactured thereafter.
  • screening methods for Effective Compounds including effective stasis compounds.
  • the methods of screening may be for oxygen antagonists or for any other compounds that can effect the methods discussed herein.
  • there are screening methods involving a) exposing a zebrafish embryo to a substance; b) measuring the heart rate of the embryo; c) comparing the heart rate of the embryo in the presence of the substance to the heart rate in the absence of the substance, wherein a reduction of heart rate, such as by 50% or more, identifies the substance as a candidate Effective Compound.
  • zebrafish embryos it is contemplated that other non-human organisms may be used as well, such as fish, frogs, flies, shrimp, or their embryos.
  • the heart rate of the embryo is measured by counting the number of heartbeats. This can be done, in some cases, by viewing the embryo under a dissecting microscope.
  • Other screening embodiments involve: a) exposing a nematode to a substance; b) assaying one or more of the following cellular respiration factors: i) core body temperature; ii) oxygen consumption; iii) motility; or, iv) carbon dioxide production; c) comparing the cellular respiration factor of the nematode in the presence of the substance to the cellular respiration factor in the absence of the substance, wherein a reduction of the characteristic identifies the substance as a candidate Effective Compound. It is specifically contemplated that motility of the nematodes is assayed in some methods of the invention.
  • the methods first involve identifying an appropriate substance to screen.
  • the substance will be a chalcogenide, reducing agent, or have the structure of Formula I or Formula IV, or any other compound discussed herein. It is further contemplated that subsequent screens can be done in organisms considered higher or more complex than those used in preliminary or initial screens.
  • one or more cellular respiration factors will be assayed in these other organisms to further evaluate a candidate compound.
  • subsequent screens involve the use of mice, rats, dogs, etc.
  • the substance in order for the substance to be considered a candidate Effective Compound (or oxygen antagonist, or stasis inducer or protective metabolic agent, or any other subset of Effective Compounds) the substance must not kill the organism or cells in the assay and the effect must be reversible (that is, the characteristic that is altered needs to resume to its level before the exposure to the substance).
  • any method of treatment can be used in the context of a preparation of a medicament for the treatment of or protection against the specified disease or condition.
  • This includes, but is not limited to, the preparation of a medicament for the treatment of hemorrhagic or hematologic shock, wounds and tissue damage, hyperthermia, hypothermia, neurodegeneration, sepsis, cancer, and trauma.
  • the invention includes, but is not limited to, the preparation of a medicament for a treatment to prevent death, shock, trauma, organ or tissue rejection, damage from cancer therapy, neurodegeneration, and wound or tissue damage.
  • organismal stasis is not any of the following states: sleep, comatose, death, anesthetized, or grand mal seizure.
  • any embodiment involving "exposing" biological matter to an Effective Compound may also be implemented so that biological matter is provided with the Effective Compound or administered the Effective Compound.
  • the term “provide” is used according to its ordinary and plain meaning: "to supply or furnish for use” (Oxford English Dictionary), which, in the case of patients, may refer to the action performed by a doctor or other medical personnel who prescribes a particular Effective Compound or administers it directly to the patient.
  • compounds of the present invention modulate HIF and can thereby affect hypoxia, ischemia, and/or stasis as well as any other condition associated with
  • HIF ⁇ stabilization as described herein such as hemorrhagic shock. Collectively, these compounds are called the Effective Compounds. Any compound as described herein may or may not be considered an Effective Compound.
  • the Effective Compounds may be represented by any one or more of the compounds of Formulas I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X, XI, carbon monoxide, chalcogenide compounds, H 2 S and other sulfur containing compounds, protective metabolic agents and/or oxygen antagonists as described herein. It is specifically contemplated that any one or more of the compounds as described herein may not be considered an Effective Compound. Subsets of the Effective Compounds are also specifically contemplated.
  • one or more Effective Compounds modulates a hypoxia-related condition. In some embodiments, one or more Effective Compounds modulates an ischemia-related condition. In some embodiments, one or more Effective Compounds modulates EPO. In other embodiments, one or more Effective Compounds induce stasis. In yet other embodiments, one or more Effective Compounds modulates hemorrhagic shock.
  • Effective Compound refers to any molecule that may modulate HIF in biological matter by, for example, altering core body temperature.
  • An Effective Compound may be a protein or fragment thereof, a small molecule, or even a nucleic acid molecule.
  • the Effective Compound identified by the present invention may be peptide, polypeptide, polynucleotide, small molecule inhibitors or any other compounds that may be designed through rational drug design starting from known inhibitors or stimulators.
  • Suitable Effective Compounds include antisense molecules, siRNAs, ribozymes, and antibodies (including single chain antibodies), each of which would be specific for the target molecule. Such compounds are described in greater detail elsewhere in this document.
  • the inventor also contemplates that other structurally similar compounds may be formulated to mimic the key portions of the structure of the Effective Compounds. Such compounds, which may include peptidomimetics of peptide modulators, may be used in the same manner as the initial Effective Compounds. Further details and embodiments of the Effective Compounds are described more fully later in this disclosure.
  • the Effective Compounds can be administered singly or in combination with various other therapeutic approaches.
  • the compound is administered with another 2-oxoglutarate dioxygenase inhibitor, wherein the two compounds have differential specificity for individual 2-oxoglutarate dioxygenase family members.
  • the two compounds may be administered at the same time as a ratio of one relative to the other or may be administered consecutively during a treatment time course, e.g., following myocardial infarction.
  • one compound specifically inhibits HIF prolyl hydroxylase activity
  • a second compound specifically inhibits procollagen prolyl 4-hydroxylase activity.
  • the compound is administered with another therapeutic agent having a different mode of action, e.g., an ACE inhibitor (ACEI), angiotensin-II receptor blocker (ARB), diuretic, and/or digoxin.
  • ACEI ACE inhibitor
  • ARB angiotensin-II receptor blocker
  • diuretic e.g., diuretic, and/or digoxin.
  • digoxin e.g., a different mode of action
  • the compound is administered with carnitine.
  • a compound of the invention inhibits one or more 2- oxoglutarate dioxygenase enzymes.
  • the compound inhibits at least two 2-oxoglutarate dioxygenase family members, e.g., HIF prolyl hydroxylase and procollagen prolyl 4-hydroxylase, with either the same specificity or with differential specificity.
  • the compound is specific for one 2- oxoglutarate dioxygenase, e.g., HIF prolyl hydroxylase, and shows little to no specificity for other family members.
  • Some embodiments of the invention comprise methods using oral and transdermal delivery mechanisms.
  • the present invention also provides an oral formulation comprising a compound of the invention.
  • the present methods involve transdermal administration of a compound of the invention.
  • the present invention also provides a transdermal patch or pad comprising a compound of the invention.
  • Another general embodiment of the present invention contemplates a method for reversibly inhibiting metabolism in an organism comprising: (i) identifying an organism in need of reversible metabolism inhibition; and;
  • Another general embodiment of the present invention contemplates a method for inducing sleep in an organism comprising:
  • Another general embodiment of the present invention contemplates a method for anesthetizing biological matter comprising: (i) identifying biological matter in need of anesthesia and;
  • Another general embodiment of the present invention contemplates a method of protecting biological matter from an injury, the onset or progression of a disease, or death comprising:
  • Another general embodiment of the present invention contemplates a method of protecting biological matter from suffering cellular damage from a disease or adverse medical condition comprising:
  • Another general embodiment of the present invention contemplates a method for treating or preventing hemorrhagic shock in an organism comprising: (i) identifying organism in need of said treatment or prevention; and
  • Another general embodiment of the present invention contemplates a method of inhibiting rejection of an organ transplant in an organism comprising: (i) identifying an organism in need of an organ transplant; and
  • hypoxia inducible factor HIF ⁇
  • the cryopreservation of the biological material comprises perfusing the biological matter with a cryoprotectant and lowering the temperature of the biological matter.
  • the biological matter or organism is provided or contacted with at least one Effective Compound in an amount and for a time sufficient to cause the biological matter or organism to enter stasis.
  • the biological matter or organism may be provided or contacted with the compound in an amount and for a time sufficient to stabilize the alpha-subunit of hypoxia inducible factor (HIF ⁇ ) in cells of the biological matter or organism.
  • HIF ⁇ hypoxia inducible factor
  • Certain embodiments of the present invention comprise incubating the biological matter or organism under hypoxic conditions and/or anoxic conditions.
  • the hypoxic and/or anoxic conditions would damage the biological matter or organism in the absence of an Effective Compound.
  • the biological matter or organism is exposed to at least one Effective Compound.
  • the biological matter or organism may be exposed to an amount of compound that reduces the rate or amount of CO 2 production by the biological matter or organism by at least about two-fold.
  • the biological matter or organism may be exposed to an amount of compound that reduces the rate or amount of oxygen consumption by the biological matter or organism by at least about two-fold.
  • the biological matter or organism is exposed to an amount of compound that decreases movement or motility by at least about 10%.
  • Some embodiments of the present invention contemplate providing a combination of Effective Compounds to the biological matter or organism.
  • Such compounds may be selected from, for example, at least one compound with a chemical formula selected from the following groups:
  • the biological matter or organism may be provided with the compound before, during, or after an injury, the onset or progression of a disease, or hemorrhaging in the biological matter or organism.
  • the injury may be from an external source.
  • the compound may be provided before the injury or before the onset or progression of the disease. In certain embodiments, the compound is not provided during or after the injury or the onset or progression of the disease.
  • the injury or disease may be associated with a reduction in metabolism or temperature of the biological matter or organism.
  • the injury may be a surgery.
  • the compound may be provided during the progression of the disease, such as thalassemia, sickle cell disease, or cystic fibrosis.
  • the biological matter or organism may be provided with the compound in an amount and for a time sufficient to reduce CO 2 evolution by at least 25%.
  • the biological matter or organism may be provided with the compound in an amount and for a time that protects the matter from damage or death resulting from the injury or the onset or progression of the disease.
  • the biological matter may be provided with the compound in an amount and for a time sufficient to increase the rate of entry of the matter into stasis following the injury or the onset or progression of the disease.
  • the biological matter or organism is provided the compound by administration to the biological matter or organism intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostaticaly, intrapleurally, intratracheally, intranasally, intrathecally, intravitreally, intravaginally, intrarectally, intratumorally, intramuscularly, intraperitoneally, intraocularly, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion, via a catheter, via a lavage, or through catheterization, immersion, absorption, or adsorption.
  • Certain embodiments of the present invention further comprise exposing the biological matter to a controlled temperature and/or pressure environment.
  • the biological matter may achieve a non-physiological core temperature.
  • the biological matter may be is exposed to a controlled temperature environment that is less than about 2O 0 C.
  • the biological matter may be exposed to the controlled temperature environment prior to and/or concurrent with being provided with one or more compounds.
  • one or more Effective Compounds comprises a cationic structure capable of targeting the one or more compounds to mitochondria.
  • the biological matter or organism is to be preserved, such as for future consumption.
  • the biological matter or organism may be used for research purposes, for example.
  • the biological matter to be preserved may include platelets, or the biological matter may be used for transplantation purposes.
  • cardioplegia is induced in the biological matter or organism.
  • the biological matter or organism is at risk for hemorrhagic shock.
  • Certain methods of the present invention further comprise monitoring the biological matter or organism for toxicity from the compound.
  • the biological matter or organism is hemorrhaging.
  • the biological matter or organism goes into hemorrhagic shock.
  • the disease or adverse medical condition may be selected from the group consisting of: hemorrhagic shock, myocardial infarction, acute coronary syndrome, cardiac arrest, neonatal hypoxia/ischemia, ischemic reperfusion injury, unstable angina, post-angioplasty, aneurysm, trauma, blood loss, an infectious disease, a hyperproliferative disease, a neurodegenerative disease, or an inflammatory disease.
  • the trauma may be surgery, stroke, heart attack, bone fracture, soft tissue damage, internal bleeding, organ damage, amputation, concussion, burns, and/or is caused by gunshot, a shrapnel wound, or a knife wound.
  • Another general embodiment of the present invention contemplates a method for stabilizing the alpha-subunit of hypoxia inducible factor (HIF ⁇ ) in biological matter, wherein the method comprises administering at least one compound with a chemical formula selected from the following groups:
  • (t) oxygen antagonist or a salt, ester, or precursor thereof, wherein the compound inhibits hydroxylation of HIF ⁇ .
  • Another general embodiment of the present invention contemplates a method for stabilizing the alpha-subunit of hypoxia inducible factor (HIF ⁇ ) in biological matter, the method comprising administering at least one compound with a chemical formula selected from the following groups:
  • the 2-oxoglutarate dioxygenase enzyme is selected from the group consisting of EGLNl, EGLN2, EGLN3, procollagen prolyl 4-hydroxylase, procollagen prolyl 3 -hydroxylase, procollagen lysyl hydroxylase, PHD4, FIH-I, and any subunit or fragment thereof.
  • Another general embodiment of the present invention contemplates a method for stabilizing the alpha-subunit of hypoxia inducible factor (HIF ⁇ ) in biological matter, the method comprising administering at least one compound with a chemical formula selected from the following groups:
  • the HIF prolyl hydroxylase enzyme is selected from the group consisting of EGLNl, EGLN2, EGLN3, and any subunit or fragment thereof.
  • Another general embodiment of the present invention contemplates a method for treating, preventing, or pretreating a HIF-associated condition in biological matter, the method comprising administration of at least one compound with a chemical formula selected from the following groups: (n) Formula (I);
  • Another general embodiment of the present invention contemplates a method for treating, preventing, or pretreating a HIF-associated condition in biological matter, the method comprising administration of at least one compound with a chemical formula selected from the following groups:
  • Another general embodiment of the present invention contemplates a method for treating, preventing, or pretreating a HIF-associated condition in biological matter, the method comprising administration of at least one compound with a chemical formula selected from the following groups:
  • methods for treating, preventing, or pretreating a HIF-associated condition in biological matter wherein the HIF-associated condition is associated with hypoxia or ischemia are contemplated.
  • the method further comprises the stabilization of HIF ⁇ , the inhibition of 2- oxoglutarate dioxygenase enzyme activity, and/or the inhibition of HIF prolyl hydroxylase enzyme activity.
  • the HIF-associated condition is associated with a pulmonary disorder, a cardiac disorder, a neurological disorder or with an ischemic event.
  • the ischemic event may be an acute ischemic event.
  • the acute ischemic event may be associated with surgery, organ transplantation, infarction, trauma, or injury.
  • the ischemic event may be a chronic ischemic event.
  • the chronic ischemic event may be associated with a condition selected from the group consisting of hypertension, diabetes, occlusive arterial disease, chronic venous insufficiency, Raynaud's disease, cirrhosis, congestive heart failure and systemic sclerosis.
  • Another general embodiment of the present invention contemplates a method of treating, preventing or pretreating a condition mediated at least in part by hypoxia inducible factor (HIF) and/or erythropoietin (EPO), said method comprising administering to a subject an effective amount of at least one compound with a chemical formula selected from the following groups:
  • HIF hypoxia inducible factor
  • EPO erythropoietin
  • (t) oxygen antagonist or a salt, ester, or precursor thereof; wherein HIF ⁇ is stabilized.
  • the condition may be selected from the group consisting of anemic disorders; neurological disorders and/or injuries; including cases of stroke, trauma, epilepsy, neurodegenerative disease, myocardial infarction, liver ischemia, renal ischemia, and stroke; peripheral vascular disorders, ulcers, burns, and chronic wounds; pulmonary embolism; and ischemic-reperfusion injury.
  • angiotensin-II receptor blocker ARB
  • any one or more of the administered compounds inhibits 2-oxoglutarate dioxygenase enzyme activity and/or HIF prolyl hydroxylase activity.
  • Certain embodiments contemplate stabilization of HIF ⁇ by specifically inhibiting hydroxylation of at least one amino acid residue in HIF ⁇ using one or more of the Effective Compounds disclosed herein.
  • one or more of the Effective Compounds are selected from the group consisting of:
  • Another general embodiment of the present invention contemplates a method for increasing expression of angiogenic factors in biological matter, the method comprising administration of at least one compound with a chemical formula selected from the following groups: (n) Formula (I);
  • Another general embodiment of the present invention contemplates a method for increasing expression of glycolytic factors in biological matter, the method comprising administration of at least one compound with a chemical formula selected from the following groups:
  • (t) oxygen antagonist or a salt, ester, or precursor thereof; wherein HIF ⁇ is stabilized.
  • Another general embodiment of the present invention contemplates a method for increasing expression of factors associated with oxidative stress in a subject, the method comprising administration of at least one compound with a chemical formula selected from the following groups: (n) Formula (I);
  • Another general embodiment of the present invention contemplates a method of treating a subject having a disorder associated with ischemic reperfusion injury, the method comprising administration of at least one compound with a chemical formula selected from the following groups:
  • HIF ⁇ is selected from the group consisting of HIF- l ⁇ , HIF -2 ⁇ , HIF-3 ⁇ , and any fragment thereof.
  • Certain methods of the present invention further comprise providing to the organism at least one compound with a chemical formula selected from the following groups: heterocyclic carboxamides; phenanthrolines; hydroxamates; or a salt, ester, or precursor thereof.
  • the heterocyclic carboxamides may be selected from the group consisting of pyridine carboxamides, quinoline carboxamides, isoquinoline carboxamines, cinnoline carboxamides and beta-carboline carboxamides.
  • the biological matter or organism utilized in one of the methods of the present invention is a cell, tissue or organ.
  • the cell, tissue, or organ may be derived from a system selected from the group consisting of the renal, cardiac, hepatic, pulmonary, hematopoietic, gastrointestinal, neuronal and musculoskeletal systems.
  • the biological matter may be an organ, tissue, or cell from the heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood, small intestine, large intestine, brain, spinal cord, smooth muscle, skeletal muscle, ovary, testis, uterus, or umbilical cord.
  • the biological matter may comprise the following cell types: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, barrier function cell, contractile cell, absorptive cell, mucosal cell, limbus cell (from cornea), stem cell, unfertilized or fertilized oocyte, or sperm.
  • the biological matter or organism may be a fly, fish, frog, or embryo thereof.
  • the biological matter or organism may be a mammal, such as a human.
  • the biological matter or organism is provided or contacted with an effective amount of the compound.
  • the effective amount may be a sublethal dose of the compound.
  • the effective amount may be a near-lethal dose of the compound.
  • Certain methods of the present invention take place in vivo.
  • compositions and methods of the invention involve a HIF-I antibody.
  • the invention relates to an antibody that specifically recognizes an epitope on HIF-I.
  • the antibody immunologically reacts with a HIF-I polypeptide, which may be from a mammal, such as a human or from a non-mammal, such as a nematode.
  • the antibody recognizes the N-terminal portion of a HIF-I polypeptide, which means it may recognize amino acids at positions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106
  • HIF-associated conditions refer to any of the ischemic, hypoxic, and other conditions as described herein, and further refer to stasis.
  • Ischemia refers to a reduction in blood flow. Ischemia is associated with a reduction in nutrients, including oxygen, delivered to tissues. Ischemia may arise due to conditions such as atherosclerosis, formation of a thrombus in an artery or vein, or blockage of an artery or vein by an embolus, vascular closure due to other causes, e.g., vascular spasm, etc. Such conditions may reduce blood flow, producing a state of hypoperfusion to an organ or tissue, or block blood flow completely. Other conditions that can produce ischemia include tissue damage due to trauma or injury, such as, e.g., spinal cord injury; viral infection, which can lead to, e.g., congestive heart failure, etc.
  • ischemic conditions and “ischemic disorders” refer to any condition, disease, or disorder that is associated with ischemia and may refer to acute ischemic conditions including, but not limited to, myocardial infarction, ischemic stroke, pulmonary embolism, perinatal hypoxia, circulatory shock including, e.g., hemorrhagic, septic, cardiogenic, etc., mountain sickness, acute respiratory failure, etc., chronic ischemic conditions including atherosclerosis, chronic venous insufficiency, chronic heart failure, cardiac cirrhosis, diabetes, macular degeneration, sleep apnea, Raynaud's disease, systemic sclerosis, nonbacterial thrombotic endocarditis, occlusive artery disease, angina pectoris, TIAs, chronic alcoholic liver disease, etc. Ischemic conditions may also result from angioplasty as well as when individuals are placed under general anesthesia, and can also cause tissue damage in organs prepared for transplant.
  • acute ischemic conditions including, but not limited
  • hypoxia refers to an environment with levels of oxygen below normal.
  • hypoxia may occur when the normal physiologic levels of oxygen are not supplied to a tissue or cell.
  • Normaloxia refers to normal physiologic levels of oxygen for the particular cell type, cell state or tissue in question.
  • Anoxia is the absence of oxygen.
  • Hypoxia may be induced in cells by culturing the cells in a reduced oxygen environment, or cells may be treated with compounds that mimic hypoxia. Determining oxygen levels that define hypoxia in cell culture is well within the skill in the art.
  • hypoxia refers to any condition, disease, or disorder that is associated with hypoxia including, but not limited to, ischemic disorders (ischemic hypoxia) such as those listed above, wherein hypoxia results from reduced circulation; pulmonary disorders (hypoxic hypoxia) such as COPD (chronic obstructive pulmonary disease), severe pneumonia, pulmonary edema, pulmonary hypertension, hyaline membrane disease, and the like, wherein hypoxia results from reduced oxygenation of the blood in the lungs; anemic disorders (anemic hypoxia) such as gastric or duodenal ulcers, liver or renal disease, thrombocytopenia or blood coagulation disorders, cancer or other chronic illness, cancer chemotherapy and other therapeutic interventions that produce anemia, and the like, wherein hypoxia results from a decreased concentration of hemoglobin or red blood cells; and altitude sickness, etc.
  • ischemic hypoxia such as those listed above, wherein hypoxia results from reduced circulation
  • pulmonary disorders such as COPD (chronic obstructive pulmonary disease), severe pneumonia,
  • disorders disorders
  • diseases disorders
  • conditions include, but are not limited to, those disorders described above.
  • stasis is an example of an HIF-associated condition.
  • HIF ⁇ refers to the alpha subunit of hypoxia inducible factor protein.
  • HIF ⁇ may be any human or other mammalian protein, or fragment thereof, including, but not limited to, human HIF- l ⁇ (Genbank Accession No. Q 16665), HIF- 2 ⁇ (Genbank Accession No. AAB41495), and HIF-3 ⁇ (Genbank Accession No. AAD22668); murine HIF-l ⁇ (Genbank Accession No. Q61221), HIF-2 ⁇ (Genbank Accession No. BAA20130 and AAB41496), and HIF-3 ⁇ (Genbank Accession No. AAC72734); rat HIF- l ⁇ (Genbank Accession No.
  • HIF ⁇ may also be any non- mammalian protein or fragment thereof, including Xenopus laevis HIF- l ⁇ (Genbank Accession No. CAB96628), Drosophila melanogaster HIF-I ⁇ (Genbank Accession No. JC4851), and chicken HIF-l ⁇ (Genbank Accession No. BAA34234).
  • HIF ⁇ gene sequences may also be obtained by routine cloning techniques, for example, by using all or part of a HIF ⁇ gene sequence described above as a probe to recover and determine the sequence of a HIF ⁇ gene in another species.
  • a fragment of HIF ⁇ includes any fragment retaining at least one functional or structural characteristic of HIF ⁇ . Fragments of HIF ⁇ include the regions defined by human HIF- l ⁇ from amino acid 401 to 603 (Huang et al, supra), amino acid 531 to 575 (Jiang et al. (1997)), amino acid 556 to 575 (Tanimoto et al, supra), amino acid 557 to 571 (Srinivas et al.
  • a fragment of HIF ⁇ includes any fragment containing at least one occurrence of the motif LXXLAP, e.g., as occurs in the HIF- l ⁇ native sequence at L 397 TLLAP and L 559 EMLAP.
  • a HIF peptide may comprise [methoxycoumarin]-DLDLEALAPYIPA-DDDFQL-amide (SEQ ID NO:5).
  • the terms "HIF prolyl hydroxylase” and "HIF PH" refer to any enzyme capable of hydroxylating a proline residue in the HIF protein.
  • the proline residue hydroxylated by HIF PH may include the proline found within the motif LXXLAP, e.g., as occurs in the human HIF- l ⁇ native sequence at L 397 TLLAP and L 559 EMLAP.
  • HIF PH includes members of the Egl-Nine (EGLN) gene family described by Taylor (2001), and characterized by Aravind and Koonin (2001), Epstein et al. (2001), and Bruick and McKnight (2001).
  • Examples of HIF PH enzymes include human SM-20 (EGLNl) (GenBank Accession No. AAG33965; Dupuy et al. (2000)), EGLN2 isoform 1 (GenBank Accession No.
  • HIF PH may include Caenorhabditis elegans EGL-9 (GenBank Accession No. AAD56365) and Drosophila melanogaster CGI 114 gene product (GenBank Accession No. AAF52050).
  • HIF PH also includes any fragment retaining at least one structural or function feature of the foregoing full-length proteins, including a fragment having hydroxylase activity. HIF PH also includes any fragment of the foregoing full-length proteins that retain at least one structural or functional characteristic.
  • amino acid sequence typically refers to HIF ⁇ and fragments thereof, or HIF PH and fragments thereof, and contemplate an oligopeptide, peptide, or protein sequence, or to a fragment of any of these, and to naturally occurring or synthetic molecules.
  • “Fragments” can refer to any portion of a sequence that retains at least one structural or functional characteristic of the protein. Immunogenic fragments or antigenic fragments are fragments of polypeptides, (e.g., fragments of about five to fifteen amino acids in length, that retain at least one biological or immunological activity.
  • amino acid sequence is used to refer to the polypeptide sequence of a naturally occurring protein molecule, “amino acid sequence” and like terms are not meant to limit the amino acid sequence to the complete native sequence associated with the recited protein molecule.
  • related proteins typically refers to, for example, proteins related to HIF ⁇ prolyl hydroxylase, and may encompass other 2-oxoglutarate dioxygenase enzymes, especially those family members that similarly require Fe , 2- oxoglutarate, and oxygen to maintain hydroxylase activity.
  • Such enzymes include, but are not limited to, e.g., procollagen lysyl hydroxylase, procollagen prolyl 4- hydroxylase, and Factor Inhibiting HIF (FIH), an asparaginyl hydroxylase responsible for regulating transactivation of HIF ⁇ .
  • procollagen lysyl hydroxylase procollagen prolyl 4- hydroxylase
  • FHI Factor Inhibiting HIF
  • Treatment and “treating” refer to administration or application of an agent, drug, or remedy to a subject or performance of a procedure or modality on a subject for the purpose of obtaining a therapeutic benefit of a disease or health-related condition.
  • a “disease” or “health-related condition” can be any pathological condition of a body part, an organ, a tissue, or a system resulting from any cause, such as infection, genetic defect, and/or environmental stress.
  • the cause may or may not be known. Examples of such conditions include, but are not limited to, premalignant states, dysplasias, cancer, and other hyperproliferative diseases as well as ischemic and hypoxic conditions and EPO-associated conditions.
  • the cancer for example, may be a recurrent cancer or a cancer that is known or suspected to be resistant to conventional therapeutic regimens and standard therapies.
  • therapeutic benefit refers to anything that promotes or enhances the well-being of the subject with respect to the medical treatment of his condition, which includes, but is not limited to, treatment of pre-cancer, dysplasia, cancer, and other hyperproliferative diseases, ischemic and hypoxic conditions and EPO-related conditions.
  • a list of nonexhaustive examples of therapeutic benefit includes extension of the subject's life by any period of time, decrease or delay in the neoplastic development of the disease, decrease in hyperproliferation, reduction in tumor growth, delay of metastases or reduction in number of metastases, reduction in cancer cell or tumor cell proliferation rate, decrease or delay in progression of neoplastic development from a premalignant condition, a decrease in pain to the subject that can be attributed to the subject's condition, an enhanced ability of biological matter to enter stasis in response to an injury or disease condition, e.g., by reducing the time or level of injury or disease required to achieve stasis, and preservation of biological matter.
  • prevention and “preventing” are used according to their ordinary and plain meaning to mean “acting before” or such an act.
  • those terms refer to administration or application of an agent, such as an Effective Compound of the present invention, drug, or remedy to a subject or performance of a procedure or modality on a subject for the purpose of blocking the onset of a disease or health-related condition.
  • an agent such as an Effective Compound of the present invention, drug, or remedy to a subject or performance of a procedure or modality on a subject for the purpose of blocking the onset of a disease or health-related condition.
  • the methods involving delivery of an Effective Compound to prevent a disease or health-related condition in a subject An amount of a pharmaceutical composition that is suitable to prevent a disease or condition is an amount that is known or suspected of blocking the onset of the disease or health- related condition.
  • anemia refers to any abnormality in hemoglobin or erythrocytes that leads to reduced oxygen levels in the blood. As such, anemia can be considered a hypoxia-related condition. Anemia can be associated with abnormal production, processing, or performance of erythrocytes and/or hemoglobin. The term anemia refers to any reduction in the number of red blood cells and/or level of hemoglobin in blood relative to normal blood levels.
  • Anemia can arise due to conditions such as acute or chronic kidney disease, infections, inflammation, cancer, irradiation, toxins, diabetes, and surgery. Infections may be due to, e.g., virus, bacteria, and/or parasites, etc. Inflammation may be due to infection, autoimmune disorders, such as rheumatoid arthritis, etc. Anemia can also be associated with blood loss due to, e.g., stomach ulcer, duodenal ulcer, hemorrhoids, cancer of the stomach or large intestine, trauma, injury, surgical procedures, etc. Anemia is further associated with radiation therapy, chemotherapy, and kidney dialysis.
  • Anemia is also associated with HIV-infected patients undergoing treatment with azidothymidine (zidovudine) or other reverse transcriptase inhibitors, and can develop in cancer patients undergoing chemotherapy, e.g., with cyclic cisplatin- or non-cisplatin-containing chemotherapeutics.
  • Aplastic anemia and myelodysplastic syndromes are diseases associated with bone marrow failure that result in decreased production of erythrocytes.
  • anemia can result from defective or abnormal hemoglobin or erythrocytes, such as in disorders including microcytic anemia, hypochromic anemia, etc.
  • Anemia can result from disorders in iron transport, processing, and utilization, see, e.g., sideroblastic anemia, etc.
  • anemic conditions and “anemic disorders” refer to any condition, disease, or disorder associated with anemia. Such disorders include, but are not limited to, those disorders listed above.
  • Anemic disorders further include, but are not limited to, aplastic anemia, autoimmune hemolytic anemia, bone marrow transplantation, Churg-Strauss syndrome, Diamond Blackfan anemia, Fanconi's anemia, Felty syndrome, graft versus host disease, hematopoietic stem cell transplantation, hemolytic uremic syndrome, myelodysplastic syndrome, nocturnal paroxysmal hemoglobinuria, osteomyelofibrosis, pancytopenia, pure red-cell aplasia, purpura Schoenlein-Henoch, sideroblastic anemia, refractory anemia with excess of blasts, rheumatoid arthritis, Shwachman syndrome, sickle cell disease, thalassemia major, thalassemia minor, thrombocytopenic purpura, etc.
  • erythropoietin-associated conditions is used inclusively and refers to any condition associated with below normal, abnormal, or inappropriate modulation of erythropoietin.
  • Erythropoietin-associated conditions include any condition wherein an increase in EPO level would provide therapeutic benefit. Levels of erythropoietin associated with such conditions can be determined by any measure accepted and utilized by those of skill in the art. Erythropoietin-associated conditions include anemic conditions such as those described above.
  • Erythropoietin-associated conditions further include neurological disorders and/or injuries, including cases of stroke, trauma, epilepsy, neurodegenerative disease and the like, wherein erythropoietin may provide a neuroprotective effect.
  • Neurodegenerative diseases contemplated by the invention include Alzheimer's disease, Parkinson's disease, Huntington's disease, and the like.
  • erythropoietin refers to any recombinant or naturally occurring erythropoietin including, e.g., human erythropoietin (GenBank Accession No. AAA52400; Lin et al. (1985)), EPOETIN human recombinant erythropoietin (Amgen, Inc., Thousand Oaks Calif), ARANESP human recombinant erythropoietin (Amgen), PROCRIT human recombinant erythropoietin (Ortho Biotech Products, L.P., Raritan N.J.), etc.
  • agonist refers to a molecule that increases or prolongs the duration of the effect of a particular molecule, e.g., an enzyme or protein, or a particular environment, e.g., hypoxia.
  • Agonists may include proteins, nucleic acids, carbohydrates, or any other molecules that modulate the effects of the target molecule.
  • Antagonist refers to a molecule which decreases the extent or duration of the effect of the biological or immunological activity of a particular molecule. Antagonists may include proteins, nucleic acids, carbohydrates, antibodies, or any other molecules that decrease the effect of the target molecule.
  • excipient means an inert or inactive substance used in the production of pharmaceutical products or other tablets, including without limitation any substance used as a binder, disintegrant, coating, compression/encapsulation aid, cream or lotion, lubricant, parenteral, sweetener or flavoring, suspending/gelling agent, or wet granulation agent.
  • Binders include, e.g., carbopol, povidone, xanthan gum, etc.; coatings include, e.g., cellulose acetate phthalate, ethylcellulose, gellan gum, maltodextrin, etc.; compression/encapsulation aids include, e.g., calcium carbonate, dextrose, fructose dc, honey dc, lactose (anhydrate or monohydrate; optionally in combination with aspartame, cellulose, or microcrystalline cellulose), starch dc, sucrose, etc.; disintegrants include, e.g., croscarmnellose sodium, gellan gum, sodium starch glycolate, etc.; creams and lotions include, e.g., maltodextrin, carrageenans, etc.; lubricants include, e.g., magnesium stearate, stearic acid, sodium stearyl fumarate, etc.; materials for chewable tablets include,
  • sweeteners include, e.g., aspartame, dextrose, fructose dc, sorbitol, sucrose dc, etc.
  • wet granulation agents include, e.g., calcium carbonate, maltodextrin, microcrystalline cellulose, etc.
  • sample is used herein in its broadest sense. Samples may be derived from any source, for example, from bodily fluids, secretions, tissues, cells, or cells in culture including, but not limited to, saliva, blood, urine, serum, plasma, vitreous, synovial fluid, cerebral spinal fluid, amniotic fluid, and organ tissue ⁇ e.g., biopsied tissue); from chromosomes, organelles, or other membranes isolated from a cell; from genomic DNA, cDNA, RNA, mRNA, etc.; and from cleared cells or tissues, or blots or imprints from such cells or tissues.
  • bodily fluids secretions, tissues, cells, or cells in culture including, but not limited to, saliva, blood, urine, serum, plasma, vitreous, synovial fluid, cerebral spinal fluid, amniotic fluid, and organ tissue ⁇ e.g., biopsied tissue
  • chromosomes, organelles, or other membranes isolated from a cell from genomic DNA, cDNA, RNA, m
  • Samples may be derived from any source, such as, for example, a human subject, or a non-human mammalian subject, etc. Also contemplated are samples derived from any animal model of disease. A sample can be in solution or can be, for example, fixed or bound to a substrate. A sample can refer to any material suitable for testing for the presence of HIF ⁇ or of fragments of HIF ⁇ or suitable for screening for molecules that bind to HIF ⁇ or to fragments thereof. Methods for obtaining such samples are within the level of skill in the art.
  • Subjects may include isolated cells, either prokaryotic or eukaryotic, or tissues grown in culture.
  • Subjects may include animals, such as a mammalian species including rat, rabbit, bovine, ovine, porcine, murine, equine, and primate, particularly human.
  • any agent or solution used with a biological sample that is living and that will be used as a living material will be pharmaceutically acceptable or pharmacologically acceptable.
  • pharmaceutically-acceptable or “pharmacologically-acceptable” refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human.
  • FIG. 1 Genetic studies of sulfide activity and tolerability in sulfide adapted C. elegans.
  • FIG. 2 Genetic studies of sulfide activity and tolerability in C. elegans.
  • the present invention generally relates to compounds and methods relating to the modulation of HIF and HIF-related conditions.
  • HIF ⁇ Stabilization of HIF ⁇ is typically modulated by the inhibition of proline hydroxylation and such HIF ⁇ stabilization is effective for treating or preventing the development or persistence of hypoxic conditions such as altitude sickness and anemia, and ischemic conditions such as deep vein thrombosis, angina pectoris, pulmonary embolism, stroke, myocardial infarction, etc.
  • the stabilization of HIF ⁇ such as via the inhibition of one or more 2-oxoglutarate dioxygenases ⁇ e.g., a HIF prolyl hydroxylase), is also associated with stasis, as described herein.
  • compounds that modulate HIF such as those of the present invention, may, in certain embodiments, treat or induce HIF-related conditions.
  • HIF-related conditions e.g., hypoxia, ischemia and stasis
  • HIF ⁇ stabilization a discussion of methods of using the Effective Compounds for the treatment or inducement of conditions associated with HIF ⁇ stabilization is also provided.
  • hypoxia the condition that induces the production of HIF ⁇
  • hypoxia is a state of reduced oxygen, which can occur when the lungs are compromised or blood flow is reduced.
  • Ischemia reduction in blood flow, can be caused by the obstruction of an artery or vein by a blood clot (thrombus) or by any foreign circulating matter (embolus), or by a vascular disorder such as atherosclerosis.
  • Reduction in blood flow can have a sudden onset and short duration (acute ischemia), or can have a slow onset with long duration or frequent recurrence (chronic ischemia).
  • Acute ischemia is often associated with regional, irreversible tissue necrosis (an infarct), whereas chronic ischemia is usually associated with transient hypoxic tissue injury.
  • Infarctions commonly occur in the spleen, kidney, lungs, brain, and heart, producing disorders such as intestinal infarction, pulmonary infarction, ischemic stroke, and myocardial infarction.
  • ischemic disorders depend on the duration and severity of ischemia, and on the length of patient survival. Necrosis can be seen within the infarct in the first 24 hours, and an acute inflammatory response develops in the viable tissue adjacent to the infarct with leukocytes migrating into the area of dead tissue. Over succeeding days, there is a gradual breakdown and removal of cells within the infarct by phagocytosis, and replacement with a collagenous or glial scar. Hypoperfusion or infarction in one organ often affects other organs. For example, ischemia of the lung, caused by, for example, a pulmonary embolism, not only affects the lung, but also puts the heart and other organs, such as the brain, under hypoxic stress.
  • Myocardial infarction which often involves coronary artery blockage due to thrombosis, arterial wall vasospasms, or viral infection of the heart, can lead to congestive heart failure and systemic hypotension. Secondary complications such as global ischemic encephalopathy can develop if the cardiac arrest is prolonged with continued hypoperfusion. Cerebral ischemia, most commonly caused by vascular occlusion due to atherosclerosis, can range in severity from transient ischemic attacks (TIAs) to cerebral infarction or stroke. While the symptoms of TIAs are temporary and reversible, TIAs tend to recur and are often followed by a stroke.
  • TIAs transient ischemic attacks
  • Occlusive arterial disease includes coronary artery disease, which can lead to myocardial infarction, and peripheral arterial disease, which can affect the abdominal aorta, its major branches, and arteries of the legs.
  • Peripheral arterial disease includes Buerger's disease, Raynaud's disease, and acrocyanosis.
  • peripheral arterial disease is commonly caused by atherosclerosis, other major causes include, e.g., diabetes, etc.
  • Complications associated with peripheral arterial disease include severe leg cramps, angina, abnormal heart rhythms, heart failure, heart attack, stroke, and kidney failure. Ischemic and hypoxic disorders are a major cause of morbidity and mortality.
  • Cardiovascular diseases cause at least 15 million deaths every year and are responsible for 30% of deaths worldwide. Among the various cardiovascular diseases, ischemic heart disease and cerebrovascular diseases cause approximately 17% of deaths. Annually, 1.3 million cases of nonfatal acute myocardial infarction are reported, making the prevalence approximately 600 per 100,000 people. Further, an estimated five million Americans suffer from venous thrombosis every year, and approximately 600,000 of these cases result in pulmonary embolism. About one-third of the pulmonary embolisms end in death, making pulmonary embolism the third most common cause of death in the United States. HIF induction results as an early response to tissue hypoxia, and thus is a critical regulator of cellular and systemic responses to low oxygen levels.
  • HIF ⁇ protein levels are elevated in most cells in response to hypoxia and HIF ⁇ is induced in vivo when animals are subjected to anemia or hypoxia. HIF ⁇ levels rise within a few hours after the onset of hypoxia and return to baseline under continued hypoxic conditions. HIF has been implicated in numerous cellular and developmental processes including cell proliferation, angiogenesis, and cell cycle arrest. HIF ⁇ has also been associated with myocardial acute ischemia and early infarction, pulmonary hypertension, and inflammation. Although HIF ⁇ has been associated with tumor growth and metastasis, there is little indication that HIF is directly involved in tumorigenesis.
  • hypoxic preconditioning in which a target organ is subjected to brief periods of hypoxia, has been shown to protect both myocardium and brain against hypoxic-ischemic injury.
  • HIF ⁇ stabilization is closely associated with ischemia and is induced by preconditioning.
  • HIF is a basic helix-loop-helix (bHLH) PAS (Per/Arnt/Sim) transcriptional activator that mediates changes in gene expression in response to changes in cellular oxygen concentration.
  • HIF is a heterodimer containing an oxygen-regulated alpha subunit (HIF ⁇ ) and a constitutively expressed beta subunit (HIF ⁇ ), also known as aryl hydrocarbon receptor nuclear transporter (ARNT).
  • HIF ⁇ subunits are rapidly degraded by a mechanism that involves ubiquitination by the von Hippel-Lindau tumor suppressor (pVHL or VHL) E3 ligase complex.
  • pVHL or VHL von Hippel-Lindau tumor suppressor
  • This mechanism involves hydroxylation of specific proline residues of the ⁇ -subunit of HIF by oxygen-dependent enzymes belonging to the Egl-9/PH superfamily of 2- oxoglutarate-dependent dioxygenases, such as Egl-9. Shen et al, 2005. This hydroxylation step increases the subunit's affinity for VHL and subsequent degradation. Under hypoxic conditions, HIF ⁇ is not degraded, and an active HIF ⁇ / ⁇ complex accumulates in the nucleus and activates the expression of several genes including glycolytic enzymes, glucose transporter (GLUT)-I, erythropoietin (EPO), and vascular endothelial growth factor (VEGF).
  • GLUT glucose transporter
  • EPO erythropoietin
  • VEGF vascular endothelial growth factor
  • the enzyme responsible for HIF ⁇ hydroxylation is a member of the 2-oxoglutarate dioxygenase family.
  • Such enzymes include, but are not limited to, procollagen lysyl hydroxylase, procollagen prolyl 3 -hydroxylase, procollagen prolyl 4-hydroxylase ⁇ (I) and ⁇ (II), thymine 7-hydroxylase, aspartyl (asparaginyl) ⁇ -hydroxylase, .epsilon.-N- trimethyllysine hydroxylase, and . gamma. -butyrobetaine hydroxylase, etc.
  • These enzymes require oxygen, Fe , 2-oxoglutarate, and ascorbic acid for their hydroxylase activity.
  • an Effective Compounds that can be used in the methods of the invention may inhibit the target 2-oxoglutarate dioxygenase enzyme family member competitively with respect to 2-oxoglutarate and noncompetitively with respect to iron.
  • the invention is directed to use of compounds to inhibit HIF ⁇ hydroxylation and thus stabilize HIF ⁇ in an oxygen-independent manner.
  • these compounds are used to produce a specific benefit in the prevention and treatment of ischemic and hypoxic conditions.
  • the compounds induce stasis.
  • the present invention demonstrates no adverse affects of treatment with respect to scar formation.
  • the present invention contemplates a "dual-therapy" approach to treatment or prevention of conditions involving ischemia or hypoxia, including ischemia or hypoxia associated with subsequent reactive fibrosis, e.g., myocardial infarction and resultant congestive heart failure.
  • the method may use one compound that inhibits more than one 2- oxoglutarate dioxygenase enzyme, e.g., HIF prolyl hydroxylase and procollagen prolyl 4-hydroxylase, with either the same specificity or with different specificities.
  • the method may use a combination of compounds wherein each compound specifically inhibits only one 2-oxoglutarate dioxygenase enzyme, e.g., one compound specifically inhibits HIF prolyl hydroxylase and a second compound specifically inhibits procollagen prolyl 4-hydroxylase.
  • a compound of the invention inhibits one or more 2- oxoglutarate dioxygenase enzymes.
  • the compound inhibits at least two 2-oxoglutarate dioxygenase family members, e.g., HIF prolyl hydroxylase and HIF asparagine-hydroxylase (FIH-I), with either the same specificity or with differential specificity.
  • the compound is specific for one 2- oxoglutarate dioxygenase, e.g., HIF prolyl hydroxylase, and shows little to no specificity for other family members.
  • the compounds can be administered in combination with various other therapeutic approaches.
  • the compound is administered with another 2-oxoglutarate dioxygenase inhibitor, wherein the two compounds have differential specificity for individual 2-oxoglutarate dioxygenase family members.
  • the two compounds may be administered at the same time as a ratio of one relative to the other. Determination of a ratio appropriate to a given course of treatment or a particular subject is within the level of skill in the art.
  • the two compounds may be administered consecutively during a treatment time course, e.g., following myocardial infarction.
  • one compound specifically inhibits HIF prolyl hydroxylase enzyme activity
  • a second compound specifically inhibits procollagen prolyl 4-hydroxylase enzyme activity.
  • one compound specifically inhibits HIF prolyl hydroxylase enzyme activity, and a second compound specifically inhibits HIF asparaginyl- hydroxylase enzyme activity.
  • the compound is administered with another therapeutic agent having a different mode of action, e.g., an ACE inhibitor (ACEI), angiotensin-II receptor blocker (ARB), statin, diuretic, digoxin, carnitine, etc.
  • ACEI ACE inhibitor
  • ARB angiotensin-II receptor blocker
  • statin diuretic, digoxin, carnitine, etc.
  • Hypoxia is a common natural stress and several well conserved responses exist that facilitate cellular adaptation to hypoxic environments. To compensate for the decrease in the capacity for aerobic energy production in hypoxia, the cell must either increase anaerobic energy production or decrease energy demand (Hochachka et al, 1996). Examples of both of these responses are common in metazoans and the particular response used depends, in general, on the amount of oxygen available to the cell.
  • hypoxia-inducible transcription factor HIF-I
  • HIF-I hypoxia-inducible transcription factor
  • anoxia defined here as ⁇ 0.001 kPa O 2 — oxidative phosphorylation ceases and thus the capacity to generate energy is drastically reduced.
  • the cell In order to survive in this environment, the cell must decrease energy demand by reducing cellular activity (Hochachka et al, 2001). For example, in turtle hepatocytes deprived of oxygen, a directed effort by the cell to limit activities such as protein synthesis, ion channel activity, and anabolic pathways results in a 94% reduction in demand for ATP (Hochachka et al, 1996).
  • C. elegans In zebrafish (Danio rerio) embryos, exposure to anoxia leads to a complete arrest of the heartbeat, movement, cell cycle progression, and developmental progression (Padilla et al., 2001). Similarly, C. elegans respond to anoxia by entering into suspended animation, in which all observable movement, including cell division and developmental progression, ceases (Padilla et al, 2002; Van Voorhies et al, 2000). C. elegans can remain suspended for 24 hours or more and, upon return to normoxia, will recover with high viability. This response allows C.
  • Endogenously produced carbon monoxide can activate signaling cascades that mitigate hypoxic damage through anti- apoptotic (Brouard et al, 2003) and anti-inflammatory (Otterbein et al, 2000) activity, and similar cytoprotective effects can be achieved in transplant models by perfusion with exogenous carbon monoxide (Otterbein et al, 2003; Amersi et al, 2002).
  • carbon monoxide competes with oxygen for binding to iron-containing proteins, such as mitochondrial cytochromes and hemoglobin (Gorman et al, 2003), though the cytoprotective effect that this activity may have in hypoxia has not been investigated.
  • hypoxia is still often a damaging stress.
  • mammals have both heme oxygenase- 1 and HIF-I, and some evidence suggests that suspended animation is possible in mammals as well (Bellamy et al, 1996; Alam et al, 2002).
  • hypoxic damage due to trauma such as heart attack, stroke or blood loss is a major cause of death.
  • the understanding of the limitations of the two fundamental strategies for surviving hypoxic stress, remaining animated or suspending animation is hampered by the fact that it has been based on studies in a variety of systems under a variety of conditions.
  • hypoxic conditions include conditions in which oxygen concentration is at or less than normal atmospheric conditions, that is less that 20.8, 20, 19, 18, 17, 16,
  • hypoxic conditions include anoxic conditions, although in some embodiments, hypoxic conditions of not less than 0.5% are implemented.
  • "normoxic conditions” constitute oxygen concentrations of around 20.8% or higher. Standard methods of achieving hypoxia or anoxia are well established and include using environmental chambers that rely on chemical catalysts to remove oxygen from the chamber.
  • Such chambers are available commercially from, for example, BD Diagnostic Systems (Sparks, MD) as GASPAK Disposable Hydrogen + Carbon Dioxide Envelopes or BIO-BAG Environmental Chambers.
  • oxygen may be depleted by exchanging the air in a chamber with a non-oxygen gas, such as nitrogen.
  • Oxygen concentration may be determined, for example using a FYRITE Oxygen Analyzer (Bacharach, Pittsburgh PA).
  • the oxygen concentration of the environment containing biological matter can be about, at least about, or at most about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,
  • a change in concentration can be any of the above percentages or ranges, in terms of a decrease or increase compared to room air or to a controlled environment.
  • the present invention provides, in certain embodiments, methods of stabilizing HIF ⁇ and/or inhibiting HIF prolyl hydroxylase, compounds that can be used in the methods, and to the use of the methods to prevent or treat disorders associated with HIF including, but not limited to, hypoxic, ischemic and/or EPO- associated disorders and uses of the methods to induce stasis.
  • the Effective Compounds comprise HIF ⁇ stabilizers and/or the HIF prolyl hydroxylase inhibitors.
  • a subset of the Effective Compounds comprising one or more Effective Compound is contemplated.
  • the present invention further relates to the discovery that stabilization of the alpha subunit of hypoxia inducible factor (HIF ⁇ ) is an effective therapeutic approach with unexpected benefits when applied to treatment or prevention of conditions associated with hypoxia and/or ischemia, e.g., myocardial infarction, stroke, occlusive arterial disease, angina pectoris, cardiac cirrhosis, atherosclerosis, shock, etc.
  • the present invention contemplates methods of stabilizing HIF ⁇ to augment angiogenesis, the response to acute hypoxia, and adaptation to chronic hypoxia.
  • the identification of methods that stabilize HIF ⁇ is beneficial in the treatment of hypoxic conditions. Further, the methods can be used to produce the beneficial effects of, e.g., a preconditioning hypoxic response, by stabilizing HIF ⁇ in a normoxic environment prior to an ischemic or hypoxic event.
  • the methods can also be used to induce HIF ⁇ - specific effects, as described below, including therapeutic angiogenesis to restore blood flow to damaged tissues; neuroprotection to prevent, e.g., apoptotic loss of neurons associated with neurodegenerative diseases; and protection against oxidative damage produced by reactive oxygen species resulting from, e.g., reperfusion following an ischemic or hypoxic event.
  • the disorder may be an acute ischemic disorder such as pulmonary, intestinal, cerebral, and/or myocardial infarction, or a chronic ischemic condition such as occlusive arterial disease, liver cirrhosis, congestive heart failure, etc.
  • the methods of the invention can be used to treat ischemia due to a transient or acute trauma, insult, or injury such as, e.g., a spinal cord injury, or to treat a patient diagnosed with, e.g. , a pulmonary disorder such as pulmonary embolism and the like.
  • the present invention provides a method for increasing expression of angiogenic factors in a subject, the method comprising stabilizing HIF ⁇ .
  • the present invention provides a method of increasing expression of glycolytic factors in a subject, the method comprising stabilizing HIF ⁇ .
  • the invention provides a method of increasing expression of factors associated with oxidative stress in a subject, the method comprising stabilizing HIF ⁇ .
  • a method of treating a subject having a disorder associated with ischemic reperfusion injury, the method comprising stabilizing HIF ⁇ is also contemplated.
  • HIF ⁇ HIF-dependent gene expression in vitro and in vivo, including genes encoding angiogenic factors such as VEGF, FIt-I, EG-VEGF, PAI-I, adrenomedullin, and Cyr ⁇ l.
  • VEGF vascular endothelial growth factor
  • FIt-I vascular endothelial growth factor
  • EG-VEGF vascular endothelial growth factor
  • PAI-I adrenomedullin
  • Cyr ⁇ l angiogenic factor
  • the ability to stabilize HIF ⁇ has potential benefits in the induction of angiogenesis and prevention of tissue damage due to ischemia and hypoxia.
  • transgenic mice expressing constitutively active HIF-I ⁇ in the epidermis show enhanced expression of each VEGF isoform and a significant increase in dermal capillaries.
  • the hypervascularity induced by HIF ⁇ shows no edema, inflammation, or vascular leakage.
  • methods of the invention can be used to induce therapeutic angiogenesis, which involves the development of collateral blood vessels to revascularize ischemic tissues. Additionally, the methods of the invention produce a dose-dependent decrease in oxygen consumption in cells without any affect on cell viability.
  • Stable HIF complexes activate expression of proteins involved in glucose uptake and utilization, such as glucose transporter (GIuT)-I and GluT-3; aldolase-A, enolase-1, hexokinase-1 and -2, and phosphofructokinase-L and -C.
  • the reduction in oxygen consumption associated with HIF ⁇ stabilization is potentially due to a shift in cellular metabolism from aerobic to anaerobic energy production.
  • the present methods can thus be applied to generate energy under low oxygen conditions, beneficial in ischemic and hypoxic conditions such as, for example, peripheral arterial disease, DVT, angina pectoris, pulmonary embolism, stroke, and myocardial infarction.
  • ischemic and hypoxic conditions such as, for example, peripheral arterial disease, DVT, angina pectoris, pulmonary embolism, stroke, and myocardial infarction.
  • Hypoxic preconditioning has been shown to effectively protect against subsequent acute ischemic insult.
  • hypoxia is stabilization of HIF ⁇ and subsequent activation of HIF -regulated genes
  • the methods of the invention will mimic hypoxic preconditioning in a normoxic environment.
  • the methods may be used prior to surgery, wherein ischemic-reperfusion injury may be expected to produce deleterious results in the patient.
  • Such preventive therapy when applied prior to an ischemic event, can be provided at any time point prior to the event, in a single or repeated dose format.
  • the methods of the invention also coordinately upregulate genes involved in oxidative stress and vascular tone.
  • genes include, e.g., inducible nitric oxide synthase (iNOS), and heme oxygenase 1.
  • iNOS inducible nitric oxide synthase
  • Production of iNOS has also been associated with the beneficial effects of hypoxic preconditioning in several animal models.
  • iNOS activity attenuates but does not abrogate the beneficial effects of preconditioning, whereas nonspecifically blocking protein production completely abrogates the benefits of preconditioning.
  • iNOS is an important component of the physiological response to preconditioning, but is not the only factor.
  • the methods of the invention coordinately regulate various factors, including iNOS, involved in hypoxic response, the methods of the invention will more accurately replicate the beneficial effects of hypoxic preconditioning.
  • the present invention provides, in certain embodiments, methods of inhibiting HIF ⁇ hydroxylation, thereby stabilizing HIF ⁇ and activating HIF- regulated gene expression.
  • the methods can be applied to the prevention, pretreatment, or treatment of conditions associated with HIF including ischemic and hypoxic conditions.
  • Such conditions include, for example, myocardial infarction, liver ischemia, renal ischemia, and stroke; peripheral vascular disorders, ulcers, burns, and chronic wounds; pulmonary embolism; and ischemic-reperfusion injury, including, for example, ischemic-reperfusion injury associated with surgery and organ transplantation.
  • the present invention provides methods of stabilizing HIF ⁇ before, during, or immediately after ischemia or hypoxia, particularly in association with myocardial infarction, stroke, or renal ischemic- reperfusion injury.
  • the invention provides methods for treating various ischemic and hypoxic conditions, in particular, using the compounds described herein.
  • the methods of the invention produce therapeutic benefit when Effective Compounds are administered following ischemia or hypoxia.
  • the Effective Compounds are selected from the group consisting of Formula I and Formula IV.
  • the methods of the invention produce a dramatic decrease in morbidity and mortality following myocardial infarction, and a significant improvement in heart architecture and performance. Further, the methods of the invention improve liver function when administered following hepatic toxic-ischemic injury. Hypoxia is a significant component of liver disease, especially in chronic liver disease associated with hepatotoxic compounds such as ethanol.
  • HIF ⁇ nitric oxide synthase and glucose transporter- 1
  • the present invention provides methods of treating conditions associated with ischemia or hypoxia, the method comprising administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, alone or in combination with a pharmaceutically acceptable excipient, to a subject.
  • the compound is administered immediately following a condition producing acute ischemia, e.g. , myocardial infarction, pulmonary embolism, intestinal infarction, ischemic stroke, and renal ischemic-reperfusion injury.
  • the compound is administered to a patient diagnosed with a condition associated with the development of chronic ischemia, e.g., cardiac cirrhosis, macular degeneration, pulmonary embolism, acute respiratory failure, neonatal respiratory distress syndrome, and congestive heart failure.
  • a condition associated with the development of chronic ischemia e.g., cardiac cirrhosis, macular degeneration, pulmonary embolism, acute respiratory failure, neonatal respiratory distress syndrome, and congestive heart failure.
  • the compound is administered immediately after a trauma or injury.
  • HIF ⁇ levels are increased by a number of factors that mimic hypoxia, including iron chelators such as desferoxamine (DFO) and divalent metal salts such as CoCl 2 HIF ⁇ levels are increased by angiotensin II, thrombin, and platelet-derived growth factor under normoxic conditions using a mechanism involving reactive oxygen species. Reports have also suggested HIF ⁇ is regulated by phosphorylation through pathways involving nitric oxide-activated phosphatidylinositol 3'-kinase (PI3K), hepatocyte growth factor, or mitogen-activated protein kinase.
  • PI3K nitric oxide-activated phosphatidylinositol 3'-kinase
  • PI3K nitric oxide-activated phosphatidylinositol 3'-kinase
  • hepatocyte growth factor or mitogen-activated protein kinase.
  • Glycogen- synthase kinase which is a downstream target of PI3K, directly phosphorylates the HIF ⁇ ODD domain.
  • Erythropoietin a naturally occurring hormone that is produced in response to HIF ⁇ , stimulates the production of red blood cells (erythrocytes), which carry oxygen throughout the body. EPO is normally secreted by the kidneys, and endogenous EPO is increased under conditions of reduced oxygen (hypoxia). All types of anemia are characterized by the blood's reduced capacity to carry oxygen, and thus are associated with similar signs and symptoms, including pallor of the skin and mucous membranes, weakness, dizziness, easy fatigability, and drowsiness, leading to a decrease in quality of life. Subjects with severe cases of anemia show difficulty in breathing and heart abnormalities. Anemia is typically associated with a condition in which the blood is deficient in red blood cells or in hemoglobin.
  • Anemia can also develop in association with chronic diseases, e.g., in inflammatory disorders, including disorders with consequent inflammatory suppression of marrow, etc.
  • Anemia may be caused by loss of blood, for example, due to accidents, surgery, or gastrointestinal bleeding caused by medications such as aspirin and ibuprofen. Excessive blood loss can also be seen in women with heavy menstrual periods, and in people with stomach ulcers, duodenal ulcers, hemorrhoids, or cancer of the stomach or large intestine, etc.
  • Various conditions can cause the destruction of erythrocytes (hemolysis), thus leading to anemia.
  • Hemolytic anemia is often caused by chemical poisoning, parasites, infection, or sickle-cell anemia.
  • the body produces antibodies against its own erythrocytes, resulting in hemolysis.
  • Any disease or injury to the bone marrow can cause anemia, since that tissue is the site of erythropoiesis, i.e. erythrocyte synthesis.
  • Irradiation, disease, or various chemical agents can also cause bone marrow destruction, producing aplastic anemia. Cancer patients undergoing chemotherapy often have aplastic anemia.
  • Anemia is also associated with renal dysfunction, the severity of the anemia correlating highly with the extent of the dysfunction. Most patients with renal failure undergoing dialysis suffer from chronic anemia. In addition to being produced in the kidney, erythropoietin is produced by astrocytes and neurons in the central nervous system (CNS), and EPO and EPO receptors are expressed at capillaries of the brain-periphery interface. Furthermore, systemically administered EPO crosses the blood-brain barrier and reduces neuronal cell loss in response to cerebral and spinal chord ischemia, mechanical trauma, epilepsy, excitotoxins, and neuroinflammation. (Sakanaka (1998); Celik et al (2002); Brines et al (2000); Calapai et al (2000); and Siren et al (2001).)
  • EPO genetically engineered EPO for the treatment of anemia in chronic renal failure patients.
  • EPO is also administered to cancer patients undergoing radiation and/or chemotherapy, decreasing the need for blood transfusions.
  • EPO is used to treat anemia associated with HIV infection or azidothymidine (AZT) therapy.
  • AZA azidothymidine
  • recombinant EPO therapy requires intravenous administration of EPO one to three times per week for up to twelve weeks, a treatment regimen that limits self- administration and is inconvenient for the patient.
  • human serum EPO shows size heterogeneity due to extensive and varied glycosylation not reproduced in any recombinant human EPO.
  • the invention includes methods that provide neuroprotective benefits, e.g., by stabilizing HIF ⁇ .
  • neuroprotective benefits e.g., by stabilizing HIF ⁇ .
  • VEGF vascular endothelial growth factor
  • EPO erythropoietin
  • EPO is a naturally occurring hormone that stimulates the production of red blood cells. See, e.g., U.S. published patent application No. 2003/0153503, incorporated herein by reference in its entirety. EPO also facilitates recovery from spinal cord injuries and provides neuroprotective benefits when induced prior to an ischemic event. (See, e.g., Gorio et al (2002); and Dawson (2002).) As the methods of the invention increase expression of neuroprotective factors such as VEGF and EPO, the methods provide neuroprotective benefit that can be applied to treatment, pretreatment, or prevention of conditions including, e.g., diabetic neuropathy, stroke, neurodegenerative disease, trauma, injury, e.g. , concussions, spinal cord injuries, etc.
  • the invention further provides methods for increasing oxygen-carrying capacity by inducing erythropoiesis, and facilitating iron transport and utilization. Specifically, certain methods of the invention increase expression of EPO via administration of one or more Effective Compounds as described herein. Methods for increasing expression of enzymes and proteins involved in iron uptake, transport, and processing are specifically contemplated. Such enzymes and proteins include, but are not limited to, transferrin and transferrin receptor, which together facilitate iron transport to and uptake by, e.g., erythroid tissue; and ceruloplasmin, a ferroxidase required to oxidize ferrous iron to ferric iron.
  • ceruloplasmin is important for supply of iron to tissues.
  • the ability of the methods of the invention to increase both endogenous erythropoietin and transport and utilization of iron provides specific advantage in oxygen delivery in both normoxic and hypoxic environments.
  • the present invention provides, in certain embodiments, methods of increasing endogenous erythropoietin (EPO). These methods can be applied in vivo, e.g., in blood plasma, or in vitro, e.g., in cell culture conditioned media.
  • the invention further provides methods of increasing endogenous EPO levels to prevent, pretreat, or treat EPO-associated conditions, including, e.g., conditions associated with anemia and neurological disorders.
  • Conditions associated with anemia include disorders such as acute or chronic kidney disease, diabetes, cancer, ulcers, infection with virus, e.g.,
  • Anemic conditions can further include those associated with procedures or treatments including, e.g., radiation therapy, chemotherapy, dialysis, and surgery.
  • Disorders associated with anemia additionally include abnormal hemoglobin and/or erythrocytes, such as found in disorders such as microcytic anemia, hypochromic anemia, aplastic anemia, etc.
  • the present methods can be used, in certain embodiments, to increase endogenous EPO in a subject undergoing a specific treatment or procedure, prophylactically or concurrently, for example, an HIV-infected anemic patient being treated with azidothymidine (zidovudine) or other reverse transcriptase inhibitors, an anemic cancer patient receiving cyclic cisplatin- or non-cisp latin-containing chemotherapeutics, or an anemic or non-anemic patient scheduled to undergo surgery.
  • Methods of increasing endogenous EPO can also be used to prevent, pretreat, or treat EPO-associated conditions associated with nerve damage or neural tissue degeneration including, but not limited to, stroke, trauma, epilepsy, spinal cord injury, and neurodegenerative disorders.
  • the methods can be used to increase endogenous EPO levels in an anemic or non-anemic patient scheduled to undergo surgery to reduce the need for allogenic blood transfusions or to facilitate banking of blood prior to surgery.
  • the small decreases in hematocrit that typically occur after presurgical autologous blood donation do not stimulate an increase in endogenous EPO or in compensatory erythropoiesis.
  • preoperative stimulation of endogenous EPO would effectively increase erythrocyte mass and autologous donation volumes while maintaining higher hematocrit levels, and such methods are specifically contemplated herein.
  • the methods of the invention could be applied to reduce allogeneic blood exposure (Crosby, 2002).
  • the methods of the invention can also be used to enhance athletic performance, improve exercise capacity, and facilitate or enhance aerobic conditioning. Such methods can be used, e.g., by athletes to facilitate training and by soldiers to improve, e.g., stamina and endurance.
  • the methods of the invention have been shown, in certain embodiments, to increase endogenous erythropoietin levels in media from cultured cells treated in vitro and in blood plasma from animals treated in vivo.
  • the kidney is the major source of erythropoietin in the body
  • other organs including brain, liver, and bone marrow, can and do synthesize erythropoietin upon appropriate stimulation.
  • endogenous erythropoietin expression can be increased in various organs of the body, including brain, kidney, and liver. Indeed, methods of the invention even increase endogenous erythropoietin levels in animals that have undergone bilateral nephrectomy.
  • the methods of the invention demonstrate, in certain embodiments, that erythropoietin levels can be increased even when kidney function is compromised.
  • the invention is not to be limited by the mechanism by which erythropoietin is produced, the decrease in erythropoietin secretion typically seen during kidney failure may be due to hyperoxia in renal tissue due to increased flowthrough/reperfusion (Priyadarshi et ah, 2002).
  • the methods of the invention increase, in certain embodiments, the hematocrit and blood hemoglobin level in animals treated in vivo.
  • the increases in plasma EPO, hematocrit, and blood hemoglobin in response to the compounds used in the methods of the invention are dose-sensitive; however, dosing regimes can be established which produce a constant, controlled level of response to the compounds of the invention. Further, treatment with compounds of the invention can correct anemia, for example, induced by a toxic compound such as the chemotherapeutic agent cisplatin, or due to blood loss, e.g., trauma, injury, parasites, or surgery.
  • the increase in hematocrit and blood hemoglobin in animals treated with an Effective Compound is preceded by an increase in the percentage of circulating immature red blood cells (reticulocytes) within the blood.
  • reticulocytes circulating immature red blood cells
  • the invention contemplates the use of the compounds of the invention in methods to increase reticulocyte levels in the blood of animals for production of cell- free reticulocyte lysates as described by, e.g., Pelham and Jackson (1976). Circulating reticulocyte levels are increased in animals, e.g., rabbits, etc., by treatment with compounds of the invention, alone or in combination with another compound such as, e.g., acetylphenylhydrazine, etc. The blood is collected, and reticulocytes are pelleted by centrifugation and lysed with distilled water. Extracts can be further processed using any appropriate methodology known to those skilled in the art. See, e.g., Jackson and Hunt (1983).
  • stasis occurs via the stabilization of HIF ⁇ , wherein one or more 2-oxoglutarate dioxygenase enzymes are inhibited.
  • one or more HIF prolyl hydroxylases are inhibited.
  • Stasis is a Latin term meaning "standstill.” In the context of stasis in living tissues, the most common forms of stasis relate to the preservation of tissues for transplant or reattachment. Typically, such tissues are immersed in a physiologic fluid, such as saline, and placed in the cold to reduce biochemical processes leading to cellular damage. This stasis is incomplete and cannot be relied upon for extended periods. In fact, the success of organ transplant and limb reattachment is inversely related to the time the organ or limb is out of contact with the intact organism.
  • a more extreme version of stasis involves placing an entire organism into what is known colloquially as "suspended animation.” Though still considered largely within the realm of science fiction, some notoriety has been achieved when wealthy individuals have sought to be cryopreserved after death, in the hope that future medical breakthroughs will permit their revival and cure of their fatal ailments. Allegedly, more than one hundred people have been cryopreserved since the first attempt in 1967, and more than one thousand people have made legal and financial arrangements for cryonics with one of several organizations, for example, Alcor Life Extension Foundation. Such methods involve the administration of anti-ischemic drugs, low temperature preservation, and methods to perfuse whole organisms with cryosuspension fluids.
  • This reflex is believed to stimulate the vagal nervous system, which controls the lungs, heart, larynx and esophagus, in order to protect vital organs.
  • cold-water stimulation of nerve receptors on the skin causes shunting of blood to the brain and to the heart, and away from the skin, the gastro-intestinal tract and extremities.
  • a protective reflex bradycardia, or slowing of the heart beat conserves the dwindling oxygen supplies within the body.
  • the expression of this reflex is not the same in all people, and is believed to be a factor in only 10-20% percent of cold-water immersion cases.
  • compositions and methods that do not rely fully or at all on hypothermia and/or oxygen may be useful in the context of organ preservation, as well as for tissue or cell preservation.
  • Cells and tissue are currently preserved using hypothermia, frequently at temperatures substantially below freezing, such as in liquid nitrogen.
  • dependence on temperature can be problematic, as apparatuses and agents for producing such low temperatures may not be readily available when needed or they may require replacement.
  • the present invention contemplates use of the Effective Compounds for the induction of reversible stasis via stabilization of HIF ⁇ , such as through the inhibition of one or more 2-oxoglutarate dioxygenase enzymes, such as HIF prolyl hydroxylase.
  • a subset of the Effective Compounds are inducers of reversible stasis.
  • compounds of Formulas I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX and/or X are contemplated.
  • Stasis a cell, tissue or organ, or organism (collectively referred to as “biological material”) is living, but cellular functions necessary for cell division, developmental progression, and/or metabolic state are slowed or even stopped. This state is desirable in a number of contexts. Stasis can be used as a method of preservation by itself, or it may be induced as part of a cryopreservation regimen. Biological materials may be preserved for research use, for transportation, for transplantation, for therapeutic treatment (such as ex vivo therapy), and to prevent the onset of trauma, for example. Stasis with respect to entire organisms has similar uses. For instance, transportation of organisms could be facilitated if they had entered stasis.
  • Stasis may be beneficial by decreasing the need of the biological material for oxygen and, therefore, bloodflow. It may extend the period of time that biological material can be isolated from a life- sustaining environment and exposed to a death-inducing environment.
  • CO 2 production is a direct marker of cellular respiration related to metabolism of an organism. This may be distinguished from "CO 2 evolution,” which refers to the amount of CO 2 blown out of the lungs.
  • Certain Effective Compounds e.g., hydrogen sulfide, can inhibit carbonic anhydrase activity in the lungs, this inhibiting conversion of carbonate to CO 2 and its liberation from the pulmonary blood, thereby exhibiting an associated reduction in CO 2 evolution, without a corresponding decrease in cellular CO 2 production.
  • the present invention is based on the observation that certain types of compounds effectively induce reversible stasis in biological matter.
  • Other patent applications discuss induction of stasis, including the following: U.S. patent applications 10/971,576, 10/972,063; and 10/971,575, all of which are hereby incorporated by reference in their entireties. a. Thermoregulation
  • thermoregulation is a characteristic of so-called "warm-blooded” animals, which permits the organism to maintain a relatively constant core body temperature even when exposed to significantly altered (cold or hot) environmental temperatures.
  • the ability to control thermoregulation by induction of stasis is one aspect of the invention, and permits uses similar to those discussed above.
  • Thermal regulation may be facilitated by placing of organisms, limbs or isolated organs or tissues into chambers/devices, the temperature of which can be controlled. For example, warm rooms or chamber-like devices similar to hyperbaric chambers may encompass an entire organism and be connected to thermo-regulatory apparti.
  • Biological matter contemplated for use with all aspects of the present invention include material derived from invertebrates and vertebrates, including mammals; biological materials includes organisms.
  • the invention can be employed with respect to mammals of veterinary or agricultural importance including those from the following classes: canine, feline, equine, bovine, ovine, murine, porcine, caprine, rodent, lagomorph, lupine, and ursine.
  • the invention also extends to fish, birds, reptiles, amphibians, invertebrates, fungi, plants, protists and prokaryotes. Such species are described in U.S. Patent Application 11/408,734, herein incorporated by reference in its entirety.
  • Stasis of the organism can be induced or stasis within cells, tissues, and/or organs of the organism can be induced.
  • Biological matter in which stasis can be induced that are contemplated for use with methods and apparatuses of the invention are limited only insofar as the comprise cells utilizing oxygen to produce energy.
  • Stasis can be induced in cells, tissues, or organs involving the heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood, small intestine, large intestine, brain, spinal cord, smooth muscle, skeletal muscle, ovary, testis, uterus, and umbilical cord.
  • stasis can be induced in cells of the following type: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, barrier function cell, contractile cell, absorptive cell, mucosal cell, limbus cell (from cornea), stem cell (totipotent, pluripotent or multipotent), unfertilized or fertilized oocyte, or sperm.
  • cells of the following type platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, barrier function cell, contractile cell, absorptive cell, mucosal cell, limbus cell (from cornea), stem cell (totipotent
  • stasis can be induced in plants or parts of plants, including fruit, flowers, leaves, stems, seeds, cuttings. Plants can be agricultural, medicinal, or decorative. Induction of stasis in plants may enhance the shelf life or pathogen resistance of the whole or part of the plant.
  • Methods and apparatuses of the invention can be used to induce stasis in in vivo biological matter. This can serve to protect and/or preserve the biological matter or the organism itself or to prevent damage or injury (or further damage or injury) to them or the organism overall. c. Trauma
  • the present invention may find use in the treatment of patients who are undergoing, or who are susceptible to trauma.
  • Trauma may be caused by external insults, such as burns, wounds, amputations, gunshot wounds, or surgical trauma, internal insults, such as stroke or heart attack that result in the acute reduction in circulation, or reductions in circulation due to non-invasive stress, such as exposure to cold or radiation. Trauma can also result from blood loss, resulting in one or more hypoxic conditions, such as EPO-related conditions ⁇ e.g., anemia), as described herein.
  • EPO-related conditions e.g., anemia
  • trauma On a cellular level, trauma often results in exposure of cells, tissues and/or organs to hypoxia, thereby resulting in induction of programmed cell death, or "apoptosis.”
  • trauma leads to the induction of a series of biochemical processes, such as clotting, inflammation, hypotension, and may give rise to shock, which if it persists may lead to organ dysfunction, irreversible cell damage and death.
  • Biological processes are designed to defend the body against traumatic insult; however they may lead to a sequence of events that proves harmful and, in some instances, fatal.
  • the present invention contemplates the placement of tissues, organs, limbs and even whole organisms into stasis as a way of protecting them from the detrimental effects of trauma.
  • one or more Effective Compounds may be administered to induce stasis via modulation of HIF (e.g., stabilization of HIF ⁇ ) and/or one or more 2-oxoglutarate dioxygenase enzymes, such as HIF prolyl hydroxylase (e.g., HIF prolyl hydroxylase inhibition); such modulation could offer physiological protection from further detrimental effects of trauma.
  • induction of stasis in vivo or ex vivo can "buy time" for the subject, either by bringing medical attention to the subject, or by transporting the subject to the medical attention.
  • the present invention also contemplates methods for inducing tissue regeneration and wound healing by prevention/delay of biological processes that may result in delayed wound healing and tissue regeneration.
  • the induction of stasis induction of stasis in vivo or ex vivo can aid in the wound healing and tissue regeneration process by managing the biological processes that inhibit healing and regeneration.
  • wounds and tissue damage are intractable or take excessive periods of time to heal. Examples are chronic open wounds (diabetic foot ulcers and stage 3 & 4 pressure ulcers), acute and traumatic wounds, flaps and grafts, and subacute wounds (i.e., dehisced incisions). This may also apply to other tissue damage, for example burns and lung damage from smoke/hot air inhalation.
  • biological matter is exposed to an Effective Compound for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more.
  • Shock is a life-threatening condition that progresses rapidly when interventions are delayed. Shock is a state in which adequate perfusion to sustain the physiologic needs of organ tissues is not present. This is a condition of profound haemodynamic and metabolic disturbance characterized by failure of the circulatory system to maintain adequate perfusion of vital organs. It may result from inadequate blood volume (hypovolemic shock), which relates to ischemia, inadequate cardiac function (cardiogenic shock) or inadequate vasomotor tone, also referred to as distributive shock (neurogenic shock, septic shock, anaphylactic shock). This often results in rapid mortality of the patient. Many conditions, including sepsis, blood loss, impaired autoregulation, and loss of autonomic tone, may produce shock or shocklike states. The present invention is anticipated to prevent detrimental effects of all the above states of shock, and sustain the life of the biological matter undergoing such shock.
  • hemorrhagic shock blood loss exceeds the body's ability to compensate and provide adequate tissue perfusion and oxygenation. This is frequently due to trauma, but may also be caused by spontaneous hemorrhage (e.g., gastrointestinal bleeding, childbirth), surgery, and other causes. Most frequently, clinical hemorrhagic shock is caused by an acute bleeding episode with a discrete precipitating event. Less commonly, hemorrhagic shock may be seen in chronic conditions with subacute blood loss.
  • Such blood loss can be considered an ischemic condition, and it is therefore contemplated that compounds of the present invention can be used to treat or manage such blood loss, wherein one or more Effective Compounds modulates HIF and/or one or more 2-oxoglutarate dioxygenases (e.g., HIF prolyl hydroxylase).
  • one or more Effective Compounds modulates HIF and/or one or more 2-oxoglutarate dioxygenases (e.g., HIF prolyl hydroxylase).
  • HIF prolyl hydroxylase e.g., HIF prolyl hydroxylase
  • Physiologic compensation mechanisms for hemorrhage include initial peripheral and mesenteric vasoconstriction to shunt blood to the central circulation. This is then augmented by a progressive tachycardia. Invasive monitoring may reveal an increased cardiac index, increased oxygen delivery (i.e., DO 2 ), and increased oxygen consumption (i.e., VO 2 ) by tissues. Lactate levels, the acid-base status, and other markers also may provide useful indicators of physiologic status. Age, medications, and comorbid factors all may affect a patient's response to hemorrhagic shock. Failure of compensatory mechanisms in hemorrhagic shock can lead to death.
  • the invention concerns inducing a whole body hibernetic state using an Effective Compound to preserve the patient's vital organs and life.
  • This will allow for transport to a controlled environment (e.g., surgery), where the initial cause of the shock can be addressed, and then the patient brought back to normal function in a controlled manner.
  • the first hour after injury referred to as the "golden hour”
  • Stabilizing the patient in this time period is the major goal, and transport to a critical care facility (e.g., emergency room, surgery, etc.) where the injury can be properly addressed.
  • an Effective Compound for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days or more, and any range or combination therein.
  • the present inventor proposes use of the present invention to treat people with extreme hypothermia, such uses involving modulation of HIF as described herein.
  • the methods and compositions of the present invention are useful for inducing hypothermia in a mammal in need of hypothermia.
  • Hypothermia can be mild, moderate or profound.
  • Mild hypothermia comprises achievement of a core body temperature of approximately between 0.1 and 5 degrees Celsius below the normal core body temperature of the mammal.
  • the normal core body temperature of a mammal is usually between 35 and 38 degrees Celsius.
  • Moderate hypothermia comprises achievement of a core body temperature of approximately between 5 and 15 degrees Celsius below the normal core body temperature of the mammal.
  • Profound hypothermia comprises achievement of a core body temperature of approximately between 15 and 37 degrees Celsius below the normal core body temperature of the mammal.
  • Mild hypothermia is known in the art to be therapeutically useful and effective in both non-human mammals and in humans.
  • the therapeutic benefit of mild hypothermia has been observed in human clinical trials in the context of out-of- hospital cardiac arrest. Exposure of humans to mild hypothermia in the context of cardiac arrest results in a survival advantage and an improved neurological outcome compared to standard of care with normothermia, or absence of mild hypothermia
  • Methods and compositions of the present invention may have advantages over other methods known in the art, including, but not limited to, packing the subject in ice, or surrounding the subject with a "cooling tent" that circulates cool air or liquid, for inducing mild, moderate, or profound hypothermia in mammals or humans.
  • the subject resists the reduction of core body temperature below normothermia and tries to generate heat by shivering.
  • Shivering, and the body heat engendered therein can have a negative impact on the achievement of mild hypothermia by, for example, slowing the rate of decrease in the core body temperature that is achieved using the standard methods of hypothermia induction. Consequently, humans subjected to therapeutic levels of hypothermia are also treated with a drug that inhibits shivering (by blocking neurotransmission at the neuromuscular junctions) (Bernard et al, 2002).
  • methods and compositions of the present invention are combined with invasive methods or medical devices known in the art to induce therapeutic hypothermia in mammals or humans.
  • invasive methods and devices include, but are not limited to, flexible probes or catheters that can be inserted into the vasculature of the subject in need of hypothermia, wherein the temperature of the catheter is adjusted to below the normal body temperature of the subject, resulting in the cooling of blood which is in contact with the catheter. The cooled blood subsequently engenders a decrease in the core body temperature of the mammal.
  • the temperature of the catheter can be modulated so as to maintain a pre- specified core body temperature.
  • Such medical devices for achieving and maintaining mild or moderate hypothermia referred to in the art as endovascular temperature therapy, are known in the art and are described for example on the World Wide Web at innercool.com and radiantmedical.com.
  • the method provides that patients with extreme hypothermia are administered or exposed to a compound of the present invention and then gradually restored to normal temperature while withdrawing, in a controlled fashion, the compound.
  • the compound buffers the biological systems within the subject so that they may be initiated gradually without shock (or harm) to the subject.
  • a subject suffering from hypothermia with be given an oral or intravenous dose of a compound of the present invention.
  • Intravenous provision may be preferred because of the potential non-responsiveness of the subject and the ability to provide a controlled dosage over a period of time.
  • the compound of the present invention may be provide in a gaseous state, for example, using a mask for inhalation or even a sealed chamber that can house the entire subject.
  • the patient will be stabilized in terms of heart rate, respiration and temperature prior to effecting any change.
  • the ambient environmental temperature will be increased, again gradually. This may be accomplished simply by removing the subject from the hypothermic conditions.
  • a more regulated increase in temperature may be effected by adding successive layers of clothing or blankets, by use of a thermal wrap with gradual increase in heat, or if possible, by placing the subject in chamber whose temperature may be gradually increased.
  • the vital signs of the subject are monitored over the course of the temperature increase.
  • the compound of the present invention is removed from the subject's environment. Both heat and treatment using a compound of the present invention are ceased at the appropriate endpoint, judged by the medical personnel monitoring the situation, but in any event at the time the subject's temperature and other vital signs return to a normal range. Continued monitoring following cessation of treatment is recommended for a period of at least 24 hrs. e. Hyperthermia
  • the technology of the present invention could be used to control whole body temperature in certain states of hyperthermia. This would likely involve administration of a compound of the present invention through inhalation or perfused into the blood supply to induce a hibernation state via modulation of HIF. It would be useful to have the patient to be in stasis for between about 6 and about 24 hours, during which time the source of the fever can be addressed.
  • a patient is exposed to a compound of the present invention for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days or more, and any range or combination therein.
  • the present invention may find use as solutions for the treatment of coronary heart disease (CHD) including a use for cardioplegia for cardiac bypass surgery (CABG).
  • CHD coronary heart disease
  • CABG cardioplegia for cardiac bypass surgery
  • CHD may be considered an ischemia- related condition, and thus it is contemplated that one or more Effective Compounds may be administered to treat CHD, wherein the Effective Compound modulates HIF (e.g., stabilization of HIF ⁇ ) and/or one or more 2-oxoglutarate dioxygenase enzymes, such as a HIF prolyl hydroxylase (e.g., inhibition of HIF prolyl hydroxylase).
  • HIF e.g., stabilization of HIF ⁇
  • 2-oxoglutarate dioxygenase enzymes such as a HIF prolyl hydroxylase (e.g., inhibition of HIF prolyl hydroxylase).
  • CHD results from atherosclerosis, a narrowing and hardening of the arteries that supply oxygen rich blood to the heart muscle.
  • the arteries harden and become narrow due to the buildup of plaque on the inner walls or linings of the arteries. Blood flow to the heart is reduced as plaque narrows the coronary arteries. This decreases the oxygen supply to the heart muscle.
  • angina which is chest pain or discomfort that happens when the heart is not getting enough blood
  • heart attack which can occur when a blood clot suddenly cuts off most or all blood supply to part of the heart and cells in the heart muscle that do not receive enough oxygen-carrying blood begin to die, potentially causing permanent damage to the heart muscle
  • heart failure which is when the heart is unable to pump blood effectively to the rest of the body
  • arrhythmias which are changes in the normal rhythm of the heartbeats.
  • CHD cardiovascular disease
  • CABG cardioplegia
  • Cardioplegic solutions are perfused through the vessels and chambers of the heart and cause its intrinsic beating to cease, while maintaining the viability of the organ. Cardioplegia (paralysis of the heart) is desirable during open-heart surgery and during the procurement, transportation, and storage of donor hearts for use in heart transplantation procedures.
  • cardioplegic techniques employed cold crystalloid solutions to initiate and maintain intraoperative cardiac arrest.
  • blood cardioplegia facilitated aerobic myocardial metabolism during the cross-clamp period and reduced anaerobic lactate production.
  • blood cardioplegia improves oxygen carrying capacity, enhanced myocardial oxygen consumption and preserved myocardial high-energy phosphate stores.
  • cardioplegic solutions often have varying amounts of potassium, magnesium, and several other minor components.
  • drugs are added to the cardioplegic solution to aid in muscle relaxation and protection from ischemia.
  • a combination of continuous retrograde along with intermittent antegrade cardioplegia reduces myocardial lactate production, preserved ATP stores, and improved metabolic recovery after cross-clamp release.
  • Tepid (29°C) cardioplegia reduces lactate and acid production during cardioplegic arrest, and improves postoperative ventricular function.
  • Cardioplegic flows of at least 200mL/min are required to washout detrimental metabolic end-products and improve ventricular function. It is abundantly clear now that future directions in cardioplegic management will involve the use of cardioplegic additives to further improve protective effects. For example, attempts have been made to harness the beneficial effects of ischemic preconditioning using adenosine. Similarly, insulin cardioplegia has been employed in order to enhance ventricular performance by stimulating early postoperative aerobic metabolism.
  • L-arginine a nitric oxide donor has been demonstrated to be beneficial in experimental studies and may represent a further option for the enhancement of intraoperative myocardial protection.
  • Future benefit of cardioplegic supplementation is likely to be observed in high-risk with poor ventricular function, for which current protective techniques are inadequate.
  • improvements in this area hold great promise for the advancement of care in this field.
  • Ischemic-reperfusion injury during cardiac bypass surgery results in poor outcomes (both morbidity and mortality), especially due to an already weakened state of the heart.
  • Myocardial ischemia results in anaerobic myocardial metabolism. The end products of anaerobic metabolism rapidly lead to acidosis, mitochondrial dysfunction, and myocyte necrosis. High-energy phosphate depletion occurs almost immediately, with a 50 percent loss of ATP stores within 10 minutes. Reduced contractility occurs within 1 to 2 minutes, with development of ischemic contracture and irreversible injury after 30 to 40 minutes of normothermic (37 0 C) ischemia.
  • Reperfusion injury is a well-known phenomenon following restoration of coronary circulation.
  • Reperfusion injury is characterized by abnormal myocardial oxidative metabolism.
  • reperfusion may produce cytotoxic oxygen free radicals.
  • oxygen free radicals play a significant role in the pathogenesis of reperfusion injury by oxidizing sarcolemmal phospholipids and thus disrupting membrane integrity.
  • Oxidized free fatty acids are released into the coronary venous blood and are a marker of myocardial membrane phospholipid peroxidation.
  • Protamine induces complement activation, which activates neutrophils.
  • Activated neutrophils and other leukocytes are an additional source of oxygen free radicals and other cytotoxic substances.
  • the present invention provides methods and compositions for inducing cardioplegia that will provide greater protection to the heart during bypass surgery.
  • the induction of cardioplegia may involve, in certain embodiments, stabilization of HIF ⁇ via inhibiting one or more 2-oxoglutarate dioxygenase enzymes, such as a HIF prolyl hydroxylase.
  • the present invention provides a cardioplegic solution comprising a compound of the present invention dissolved in solution or bubbled as a gas in the solution.
  • the invention further comprises at least a first device, such as a catheter or cannula, for introducing an appropriate dose of the cardioplegic solution to the heart.
  • the invention further comprises at least a second device, such as a catheter or cannula, for removing the cardioplegic solution from the heart.
  • a second device such as a catheter or cannula
  • the heart is exposed to a compound of the present invention for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 hours or more, and any range or combination therein.
  • Cancer is a leading cause of mortality in industrialized countries around the world.
  • the most conventional approach to the treatment of cancer is by administering a cytotoxic agent to the cancer patient (or treatment ex vivo of a tissue) such that the agent has a more lethal effect on the cancer cells than normal cells.
  • chemo- and radiotherapy are often characterized by severe side effects, some of which are life threatening, e.g., sores in the mouth, difficulty swallowing, dry mouth, nausea, diarrhea, vomiting, fatigue, bleeding, hair loss and infection, skin irritation and loss of energy (Curran, 1998; Brizel, 1998).
  • hypothermia transient and reversible lowering of the core body temperature, or "hypothermia,” may lead to improvements in the fight against cancer.
  • Hypothermia of 28 0 C was recently found to reduce radiation, doxorubicin-and cisplatin-induced toxicity in mice.
  • the cancer fighting activity of these drugs/treatments was not compromised when administered to cooled animals; rather, it was enhanced, particularly for cisp latin (Lundgren-Eriksson et ah, 2001). Based on this and other published work, the inventor proposes a further reduction in core temperature will provide benefit to cancer patients.
  • the present invention contemplates the use of a compound of the present invention to induce stasis in normal tissues of a cancer patient, thereby reducing the potential impact of chemo- or radiotherapy on those tissues. It also permits the use of higher doses of chemo- and radiotherapy, thereby increasing the anti-cancer effects of these treatments.
  • hyperproliferative disorder including benign and malignant neoplasias, non-neoplastic hyperproliferative conditions, pre -neoplastic conditions, and precancerous lesions.
  • disorders include restenosis, cancer, multi-drug resistant cancer, primary psoriasis and metastatic tumors, angiogenesis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, eczema, and secondary cataracts, as well as oral hairy leukoplasia, bronchial dysplasia, carcinomas in situ, and intraepithelial hyperplasia.
  • the present invention is directed at the treatment of human cancers including cancers of the prostate, lung, brain, skin, liver, breast, lymphoid system, stomach, testicles, ovaries, pancreas, bone, bone marrow, gastrointestine, head and neck, cervix, esophagus, eye, gall bladder, kidney, adrenal glands, heart, colon and blood.
  • cancers involving epithelial and endothelial cells are also contemplated for treatment.
  • chemo- and radiotherapy are designed to reduce tumor size, reduce tumor cell growth, induce apoptosis in tumor cells, reduce tumor vasculature, reduce or prevent metastasis, reduce tumor growth rate, accelerate tumor cell death, and kill tumor cells.
  • compositions comprising a compound of the present invention with secondary anti-cancer agents (secondary agents) effective in the treatment of hyperproliferative disease.
  • An "anti-cancer” agent is capable of negatively affecting cancer in a subject, for example, by killing cancer cells, inducing apoptosis in cancer cells, reducing the growth rate of cancer cells, reducing the incidence or number of metastases, reducing tumor size, inhibiting tumor growth, reducing the blood supply to a tumor or cancer cells, promoting an immune response against cancer cells or a tumor, preventing or inhibiting the progression of cancer, or increasing the lifespan of a subject with cancer.
  • Secondary anti-cancer agents include biological agents (biotherapy), chemotherapy agents, and radiotherapy agents. More generally, these other compositions are provided in a combined amount effective to kill or inhibit proliferation of the cancer or tumor cells, while at the same time reducing or minimizing the impact of the secondary agents on normal cells.
  • This process may involve contacting or exposing the cells with a compound of the present invention and the secondary agent(s) at the same time. This may be achieved by contacting the cell with a single composition or pharmacological formulation that includes both agents, or by contacting or exposing the cell with two distinct compositions or formulations, at the same time, wherein one composition includes a compound of the present invention and the other includes the second agent(s).
  • the therapy comprising use of a compound of the present invention may precede or follow the secondary agent treatment by intervals ranging from minutes to weeks.
  • the other agent and expression construct are applied separately to the cell, one would generally ensure that a significant period of time did not expire between the time of each delivery, such that the agent and expression construct would still be able to exert an advantageously combined effect on the cell.
  • biological matter will be kept in stasis for between about 2 and about 4 hours while the cancer treatment is being administered.
  • biological matter is exposed to a compound of the present invention for about, at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6 hours or more, and any range or combination therein.
  • the Effective Compound is "A”
  • the secondary anti-cancer agent such as radio- or chemotherapy
  • Cancer therapies also include a variety of combination therapies with both chemical and radiation based treatments.
  • Combination chemotherapies include, for example, cisplatin (CDDP), carboplatin, procarbazine, mechlorethamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosurea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicomycin, mitomycin, etoposide (VP 16), tamoxifen, raloxifene, estrogen receptor binding agents, taxol, gemcitabien, navelbine, farnesyl-protein transferase inhibitors, transplatinum, 5-fluorouracil, vincristine, vinblastine and methotrexate, Temazolomide (an aqueous form of DTIC), or any analog or derivative variant of the foregoing.
  • Dosage ranges for X-rays range from daily doses of 50 to 200 roentgens for prolonged periods of time (3 to 4 wk), to single doses of 2000 to 6000 roentgens.
  • Dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the uptake by the neoplastic cells.
  • contacted and “exposed,” when applied to a cell are used herein to describe the process by which a composition of the invention or a chemotherapeutic or radiotherapeutic agent is delivered to a target cell or are placed in direct juxtaposition with the target cell.
  • both agents may be delivered to a cell in a combined amount effective to kill the cell or prevent it from dividing.
  • Immunotherapeutics generally, rely on the use of immune effector cells and molecules to target and destroy cancer cells.
  • the immune effector may be, for example, an antibody specific for some marker on the surface of a tumor cell.
  • the antibody alone may serve as an effector of therapy or it may recruit other cells to actually effect cell killing.
  • the antibody also may be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.) and serve merely as a targeting agent.
  • the effector may be a lymphocyte carrying a surface molecule that interacts, either directly or indirectly, with a tumor cell target.
  • Various effector cells include cytotoxic T cells and NK cells.
  • Immunotherapy could also be used as part of a combined therapy.
  • the general approach for combined therapy is discussed below.
  • the tumor cell must bear some marker that is amenable to targeting, i.e., is not present on the majority of other cells.
  • Common tumor markers include carcinoembryonic antigen, prostate specific antigen, urinary tumor associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72, HMFG, Sialyl Lewis Antigen, MucA, MucB, PLAP, estrogen receptor, laminin receptor, erb B and pi 55.
  • Immune stimulating molecules also exist including: cytokines such as IL-2, IL-4, IL-12, GM-CSF, gamma-IFN, chemokines such as MIP-I, MCP-I, IL-8 and growth factors such as FLT3 ligand.
  • cytokines such as IL-2, IL-4, IL-12, GM-CSF, gamma-IFN, chemokines such as MIP-I, MCP-I, IL-8 and growth factors such as FLT3 ligand.
  • chemokines such as MIP-I, MCP-I, IL-8
  • growth factors such as FLT3 ligand.
  • immune adjuvants ⁇ e.g., Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene and aromatic compounds
  • U.S. Patent 5,801,005 U.S.
  • cytokine therapy ⁇ e.g., interferons ⁇ , ⁇ and ⁇ ; IL-I, GM-CSF and TNF
  • gene therapy ⁇ e.g., TNF, IL- 1, IL-2, p53
  • Patent 5,846,945 and monoclonal antibodies ⁇ e.g., anti- ganglioside GM2, anti-HER-2, anti-pl85) (Pietras et al, 1998; Hanibuchi et al, 1998).
  • Herceptin tacuzumab
  • Herceptin is a chimeric (mouse-human) monoclonal antibody that blocks the HER2-neu receptor. It possesses anti-tumor activity and has been approved for use in the treatment of malignant tumors (Dillman, 1999). Combination therapy of cancer with herceptin and chemotherapy has been shown to be more effective than the individual therapies. Thus, it is contemplated that one or more anti- cancer therapies may be employed with the anti-tumor therapies described herein.
  • the present invention may be used to treat neurodegenerative diseases.
  • Neurodegenerative diseases are characterized by degeneration of neuronal tissue, and are often accompanied by loss of memory, loss of motor function, and dementia. With dementing diseases, intellectual and higher integrative cognitive faculties become more and more impaired over time. It is estimated that approximately 15% of people 65 years or older are mildly to moderately demented.
  • Neurodegenerative diseases include Parkinson's disease; primary neurodegenerative disease; Huntington's Chorea; stroke and other hypoxic or ischemic processes; neurotrauma; metabolically induced neurological damage; sequelae from cerebral seizures; hemorrhagic shock; secondary neurodegenerative disease (metabolic or toxic); Alzheimer's disease, other memory disorders; or vascular dementia, multi-infarct dementia, Lewy body dementia, or neurodegenerative dementia.
  • oxidative and reductive states which are intimately linked to circadian rhythms. That is, oxidative stress placed upon the body during consciousness is cycled to a reductive environment during sleep. This is thought to be a large part of why sleep is so important to health.
  • Certain neurodegenerative disease states such as Huntington's disease and Alzheimer's disease, as well as the normal processes of aging have been linked to a discord in this cycling pattern.
  • brain H 2 S levels are reduced in these conditions (Eto et al, 2002).
  • the present invention can be used to regulate and control the cycling between the oxidative and reduced states, for example, to prevent or reverse the effects of neurodegenerative diseases and processes.
  • Controlling circadian rhythms can have other applications, for example, to adjust these cycling patterns after traveling from one time zone to another, so as to adjust to the new time zone.
  • reduced metabolic activity overall has been shown to correlate with health in aged animals and humans. Therefore, the present invention would also be useful to suppress overall metabolic function to increase longevity and health in old age. It is contemplated that this type of treatment would likely be administered at night, during sleep for period of approximately 6 to 10 hours each day. This could require daily treatment for extended periods of time from months to years. i. Aging
  • aging itself may be thoroughly or completely inhibited for the period of time when the biological matter is in that state.
  • the present invention may inhibit aging of biological material, with respect to extending the amount of time the biological material would normally survive and/or with respect to progression from one developmental stage of life to another.
  • compositions and methods of the invention may be addressed using compositions and methods of the invention. These diseases include, but are not limited to, thalassemia and sickle cell anemia.
  • Normal hemoglobin contains two alpha and two beta globin polypeptide (protein) chains, each bound to an iron containing heme ring.
  • Thalassemia is a group of conditions in which there is an imbalance of alpha and beta chains leading to the unpaired chains precipitating on the normally fragile red blood cell membrane, leading to cell destruction. This leads to severe anemia that the marrow tries to compensate for by trying to make more red cells.
  • This process is very inefficient leading to massive expansion of the marrow space and spread of blood making to other parts of the body. This and the anemia lead to major toxicities.
  • HbA Sickle Cell Disease Normal hemoglobin
  • SCD sickle cell disease
  • HbS can polymerize (crystallize) and precipitate damaging the normally fragile red blood cell membrane, leading to cell destruction and anemia low red blood cells (RBC).
  • RBC red blood cells
  • cells with polymerized HbS change shape (sickle) and become sticky and activate mechanisms leading to coagulation and blockage of blood flow.
  • HIF prolyl hydroxylase Other conditions or events associated with HIF activity that may be modulated by the Effective Compounds via stabilization of HIF ⁇ and/or inhibition of one or more 2-oxoglutarate dioxygenase enzymes, such as HIF prolyl hydroxylase, include, but are not limited to: hypertension; diabetes; chronic venous insufficiency; Raynaud's disease; cirrhosis (including, e.g., cardiac cirrhosis); systemic sclerosis; chronic skin ulcers; formation of thrombus; vascular closure; viral infection; ischemic stroke; prenatal hypoxia; circulatory shock; altitude or mountain sickness; acute respiratory failure; nonbacterial thrombus endocarditis; chronic heart failure; macular degeneration; angina pectoris; TIAs (transient ischemic attack); chronic alcoholic liver disease; COPD (chronic obstructive pulmonary disease); severe pneumonia; pulmonary edema; pulmonary hypertension; ulcers (including, e.g
  • a further condition comprises anemia caused by or associated with acute kidney disease, chronic kidney disease, infections, inflammation (wherein the inflammation may be due to, for example, infection, autoimmune disorders, such as rheumatoid arthritis), cancer, irradiation, toxins, diabetes, surgery, virus (such as, e.g., HIV), bacteria and parasites, blood loss due to, e.g., stomach ulcer, duodenal ulcer, hemorrhoids, stomach or intestinal cancer, trauma, injury, surgical procedures, radiation therapy, chemotherapy, kidney dialysis, HIV-infected patients undergoing treatment with azidothymidine (zidovudine) or other reverse transcriptase inhibitors, and can develop in patients undergoing chemotherapy, e.g., with cyclic cisplatin- or non-cisplatin-containing chemotherapeutics, aplastic anemia, defective or abnormal hemoglobin or erythrocytes, such as in disorders including microcytic anemia and hypochromic anemia, disorders in iron transport, processing
  • an Effective Compounds may also be used in methods for treating a patient at risk of developing an ischemic or hypoxic condition, e.g., individuals at high risk for atherosclerosis, etc.
  • Risk factors for atherosclerosis include, e.g., hyperlipidemia, cigarette smoking, hypertension, diabetes mellitus, hyperinsulinemia, and abdominal obesity. Therefore, the present invention provides methods of preventing ischemic tissue injury, the method comprising administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, alone or in combination with a pharmaceutically acceptable excipient, to a patient in need.
  • the compound can be administered based on predisposing conditions, e.g., hypertension, diabetes, occlusive arterial disease, chronic venous insufficiency, Raynaud's disease, chronic skin ulcers, cirrhosis, congestive heart failure, and systemic sclerosis.
  • predisposing conditions e.g., hypertension, diabetes, occlusive arterial disease, chronic venous insufficiency, Raynaud's disease, chronic skin ulcers, cirrhosis, congestive heart failure, and systemic sclerosis.
  • compounds of the present invention modulate HIF and can thereby affect hypoxia, ischemia, and/or stasis as well as any other condition associated with HIF ⁇ stabilization as described herein, such as EPO-related conditions and hemorrhagic shock.
  • these compounds are called the Effective Compounds.
  • Subsets of the Effective Compounds for various applications are specifically contemplated.
  • the Effective Compounds or a subset thereof stabilize HIF ⁇ via interaction with a 2-oxoglutarate dioxygenase, such as HIF prolyl hydroxylase (e.g., Egl-9).
  • the Effective Compounds or a subset thereof modulate EPO.
  • any Effective Compound previously discovered to stabilize HIF ⁇ can be co-administered with any compound previously discovered to induce stasis.
  • one or more subsets of the Effective Compounds induce stasis.
  • one or more subsets of the Effective Compounds modulates hemorrhagic shock.
  • the Effective Compounds may be represented by any of Formulas I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X, XI, carbon monoxide, chalcogenide compounds, H 2 S and other sulfur containing compounds, protective metabolic agents or oxygen antagonists as described herein.
  • any subset of any of the Effective Compounds described herein is envisioned.
  • an Effective Compound may be described by one or more categories, e.g., a chalcogenide compound.
  • a particular Effective Compound may fall into more than one descriptive category, e.g., an Effective Compound may be both a chalcogenide and a compound of Formula XI.
  • an Effective Compound has a chemical structure as set forth as Formula I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X or XI described herein, or is a precursor of Formula I, Ia-Id, II, III, Ilia, IV, V, VI, VII, VIII, IX, X or XI.
  • one embodiment contemplates the subset comprising those structures represented by Formulas I and IV.
  • subset comprising those structures represented by Formulas Ia-Id, II, III, Ilia, V, VI, VII, VIII, IX, X and XL
  • subset may comprise chalcogenides as described herein.
  • subset may comprise oxygen antagonists as described herein.
  • Alkyl where used, either alone or within other terms such as “arylalkyl”, “aminoalkyl”, “thioalkyl” “cyanoalkyl” and “hydroxyalkyl”, may refer to linear or branched radicals having one to about twenty carbon atoms.
  • the term “lower alkyl” may refer to C 1 -C 6 alkyl radicals.
  • alkyl typically includes those radicals that are substituted with groups such as hydroxy, halo (such as F, Cl,
  • haloalkyl alkoxy, haloalkoxy, alkylthio, cyano, isocyano, carboxy (-COOH), alkoxycarbonyl, (-COOR), acyl, acyloxy, amino, alkylamino, urea (--NHCONHR), thiol, alkylthio, sulfoxy, sulfonyl, arylsulfonyl, alkylsulfonyl, sulfonamido, arylsulfonamido, heteroaryl, heterocyclyl, heterocycloalkyl, amidyl, alkylimino carbonyl, amidino, guanidino, hydrazino, hydrazide, sodium sulfonyl (-SOsNa), sodium sulfonylalkyl (-R SOsNa) or any combination thereof.
  • radicals may include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n- butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl and the like.
  • Hydroxyalkyl may refer to an alkyl radical, as defined herein, substituted with one or more hydroxyl radicals.
  • hydroxyalkyl radicals may include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3- hydroxypropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2,3- dihydroxypropyl, l-(hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4- dihydroxybutyl, and 2-(hydroxymethyl)-3-hydroxypropyl, and the like.
  • Arylalkyl may refer to the radical RR- wherein an alkyl radical, "R” is substituted with an aryl radical “R'.”
  • arylalkyl radicals may include, but are not limited to, benzyl, phenylethyl, 3-phenylpropyl, and the like.
  • Aminoalkyl may refer to the radical H 2 NR'-, wherein an alkyl radical is substituted with an amino radical. Examples of such radicals include may aminomethyl, aminoethyl, and the like. "Alkylaminoalkyl” may refer to an alkyl radical substituted with an alkylamino radical.
  • Alkylsulfonamido may refer to a sulfonamido group (-S(O) 2 -NRR') appended to an alkyl group, as defined herein.
  • Thioalkyl may refer to an alkyl radical substituted with one or more thiol radicals.
  • Alkylthioalkyl may refer to wherein an alkyl radical is substituted with one or more alkylthio radicals. Examples may include, but are not limited to, methylthiomethyl, ethylthioisopropyl, and the like.
  • Arylthio alkyl may refer to wherein an alkyl radical, as herein defined, is substituted with one or more arylthio radicals.
  • Carboxyalkyl may refer to the radicals -RCO 2 H, wherein an alkyl radical is substituted with a carboxyl radical. Examples may include, but are not limited to, carboxymethyl, carboxyethyl, carboxypropyl, and the like.
  • Alkylene may refer to bridging alkyl radicals.
  • alkenyl may refer to an unsaturated, acyclic hydrocarbon radical in so much as it contains at least one double bond. Such alkenyl radicals typically contain from about 2 to about 20 carbon atoms.
  • lower alkenyl may refer to C 1 -C 6 alkenyl radicals.
  • alkenyl radicals typically includes those radicals substituted as for alkyl radicals.
  • alkenyl radicals may include propenyl, 2-chloropropenyl, buten-1-yl, isobutenyl, pent-1-en-l-yl, 2-2- methy-1-buten-l-yl, 3 -methyl- 1 -buten-1-yl, hex-2-en-l-yl, 3-hydroxyhex-l-en-l-yl, hept-1-en-l-yl, and oct-1-en-l-yl, and the like.
  • alkynyl may refer to an unsaturated, acyclic hydrocarbon radical in so much as it contains one or more triple bonds, such radicals typically containing about 2 to about 20 carbon atoms.
  • lower alkynyl may refer to C 1 -C 6 alkynyl radicals.
  • alkynyl radicals typically includes those radicals substituted as for alkyl radicals.
  • alkynyl radicals may include ethynyl, propynyl, hydroxypropynyl, but-1-yn-l-yl, but-l-yn-2-yl, pent-1-yn- 1-yl, pent-l-yn-2-yl, 4-methoxypent-l-yn-2-yl, 3-methylbut-l-yn-l-yl, hex-1-yn-l-yl, hex-l-yn-2-yl, hex-l-yn-3-yl, 3, 3 -dimethyl- 1-butyn-l-yl radicals and the like.
  • Alkoxy may refer to the radical RO-, wherein R' is an alkyl radical as defined herein. Examples may include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, isopropoxy, tert-butoxy alkyls, and the like.
  • Alkoxyalkyl may refer to alkyl radicals substituted by one or more alkoxy radicals. Examples may include, but are not limited to, methoxymethyl, ethoxyethyl, methoxyethyl, isopropoxyethyl, and the like.
  • Alkoxycarbonyl may refer to the radical R-O-C(O)-, wherein R is an alkyl radical as defined herein.
  • alkoxycarbonyl radicals may include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, sec-butoxycarbonyl, isopropoxycarbonyl, and the like.
  • Alkoxythiocarbonyl may refer to R-O-C(S)-.
  • Aryl may refer to the monovalent aromatic carbocyclic radical consisting of one individual ring, or one or more fused rings in which at least one ring is aromatic in nature, which can optionally be substituted with one or more, such as one or two, substituents such as hydroxy, halo (such as F, Cl, Br, I), haloalkyl, alkoxy, haloalkoxy, alkylthio, cyano, carboxy (-COOH), alkoxycarbonyl, (-COOR), acyl, acyloxy, amino, alkylamino, urea (--NHCONHR), thiol, alkylthio, sulfoxy, sulfonyl, arylsulfonyl, alkylsulfonyl, sulfonamido, arylsulfonamido, heteroaryl, heterocyclyl, heterocycloalkyl, amidyl, alkylimino carbonyl, amidino,
  • aryl radicals may include, but are not limited to, phenyl, naphthyl, biphenyl, indanyl, anthraquinolyl, tert-butyl -phenyl, 1,3-benzodioxolyl, and the like.
  • Arylsulfonamido may refer to a sulfonamido group, as defined herein, appended to an aryl group, as defined herein.
  • Thioaryl may refer to an aryl group substituted with one or more thiol radicals.
  • Alkylamino may refer to amino groups that are substituted with one or two alkyl radicals. Examples may include monosubstituted N-alkylamino radicals and N,N-dialkylamino radicals. Other examples may include N-methylamino, N- ethylamino, N, N-dimeythylamino N,N-diethylamino, N-methyl, N-ethyl-amino, and the like.
  • Aminocarbonyl may refer to the radical H 2 NCO-.
  • Aminocarbonyalkyl may refer to the substitution of an alkyl radical, as herein defined, by one or more aminocarbonyl radicals.
  • “Amidyl” may refer to RCO-NH-, wherein R is a H or alkyl, aryl, or heteroaryl, as defined herein.
  • alkyliminocarbonyl may refer to an imino radical substituted with an alkyl group.
  • amino may refer to a substituted or unsubstituted amino group bonded to one of two available bonds of an iminocarbonyl radical.
  • guanidino may refer to an amidino group bonded to an amino group as defined above where said amino group can be bonded to a third group. Examples of such guanidino radicals include, for example, NH 2 -C(NH) -NH-, NH 2 -C(NCH 3 )-NH-, NH 2 -C(NOCH 3 )-NH-, and CH 3 NH-C(NOH)-NH-.
  • hydrazino may refer to -NH-NRR', where R and R are independently hydrogen, alkyl and the like.
  • heterocyclyl may refer to saturated and partially saturated heteroatom-containing ring-shaped radicals having from 4 through 15 ring members, herein referred to as "C 4 -C 1 S heterocyclyl” selected from carbon, nitrogen, sulfur and oxygen, wherein at least one ring atom is a heteroatom.
  • Heterocyclyl radicals may contain one, two or three rings wherein such rings may be attached in a pendant manner or may be fused.
  • saturated heterocyclic radicals may include saturated 3 to 6-membered heteromonocyclic group containing 1 to 4 nitrogen atoms
  • partially saturated heterocyclyl radicals may include dihydrothiophene, dihydropyran, dihydrofuran and dihydrothiazole.
  • Non-limiting examples of heterocyclic radicals may include 2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl, 1,3-dioxolanyl, 2H-pyranyl, 4H-pyranyl, piperidinyl, 1 ,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl, and the like.
  • heterocyclyl groups may be optionally substituted with groups such as substituents such as hydroxy, halo (such as F, Cl, Br, I), haloalkyl, alkoxy, haloalkoxy, alkylthio, cyano, carboxy (-COOH), alkoxycarbonyl, (-COOR), acyl, acyloxy, amino, alkylamino, urea (-- NHCONHR), thiol, alkylthio, sulfoxy, sulfonyl, arylsulfonyl, alkylsulfonyl, sulfonamido, arylsulfonamido, heteroaryl, heterocyclyl, heterocycloalkyl, amidyl, alkylimino carbonyl, amidino, guanidino, hydrazino, hydrazide, sodium sulfonyl (-SO 3 Na), sodium sulfonylalkyl (
  • Heteroaryl may refer to monovalent aromatic cyclic radicals having one or more rings, such as one to three rings, of four to eight atoms per ring, incorporating one or more heteroatoms, such as one or two, within the ring (chosen from nitrogen, oxygen, or sulfur), which can optionally be substituted with one or more, such as one or two substituents selected from substituents such as hydroxy, halo (such as F, Cl, Br, I), haloalkyl, alkoxy, haloalkoxy, alkylthio, cyano, carboxy (-COOH), alkoxycarbonyl, (-COOR), acyl, acyloxy, amino, alkylamino, urea (--NHCONHR), thiol, alkylthio, sulfoxy, sulfonyl, arylsulfonyl, alkylsulfonyl, sulfonamido, arylsulfonamido,
  • heteroaryl radicals may include, but are not limited to, imidazolyl, oxazolyl, thiazolyl, pyrazinyl, thienyl, furanyl, pyridinyl, quinolinyl, isoquinolinyl, benzofuryl, benzothiophenyl, benzothiopyranyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzopyranyl, indazolyl, indolyl, isoindolyl, quinolinyl, isoquinolinyl, naphthyridinyl, benzenesulfonyl-thiophenyl, and the like.
  • Heteroaryloxy may refer to heteroaryl radicals attached to an oxy radical.
  • examples of such radicals may include, but are not limited to, 2-thiophenyloxy, 2- pyrimidyloxy, 2-pyridyloxy, 3-pyridyloxy, 4-pyridyloxy, and the like.
  • Heteroaryloxyalkyl may refer to alkyl radicals substituted with one or more heteroaryloxy radicals. Examples of such radicals may include 2-pyridyloxymethyl, 3-pyridyloxyethyl, 4-pyridyloxymethyl, and the like.
  • Cycloalkyl may refer to monovalent saturated carbocyclic radicals consisting of one or more rings, typically one or two rings, of three to eight carbons per ring, which can typically be substituted with one or more substitutents, such as hydroxy, halo (such as F, Cl, Br, I), haloalkyl, alkoxy, haloalkoxy, alkylthio, cyano, carboxy (-COOH), alkoxycarbonyl, (-COOR), acyl, acyloxy, amino, alkylamino, urea (--NHCONHR), thiol, alkylthio, sulfoxy, sulfonyl, arylsulfonyl, alkylsulfonyl, sulfonamido, arylsulfonamido, heteroaryl, heterocyclyl, heterocycloalkyl, amidyl, alkylimino carbonyl, amidino, guanidino
  • cycloalkyl radicals may include, but are not limited to, cyclopropyl, cyclobutyl, 3- ethylcyclobutyl, cyclopentyl, cycloheptyl, and the like.
  • Cycloalkenyl may refer to radicals having three to ten carbon atoms and one or more carbon-carbon double bonds. Typical cycloalkenyl radicals have three to seven carbon atoms.
  • Cycloalkenylalkyl may refer to radicals wherein an alkyl radical, as defined herein, is substituted by one or more cycloalkenyl radicals.
  • Cycloalkoxy may refer to cycloalkyl radicals attached to an oxy radical. Examples may include, but are not limited to, cyclohexoxy, cyclopentoxy, and the like. “Cycloalkoxyalkyl” may refer to alkyl radicals substituted one or more cycloalkoxy radicals. Examples may include cyclohexoxyethyl, cyclopentoxymethyl, and the like.
  • Sulfmyl may refer to -S(O) -.
  • “Sulfonyl” may refer to -S(O) 2 -, wherein “alkylsulfonyl” may refer to a sulfonyl radical substituted with an alkyl radical, RSO 2 -, and “arylsulfonyl” may refer to aryl radicals attached to a sulfonyl radical. “Sulfonamido” may refer to -S(O) 2 -NRR'.
  • Sulfonic acid may refer to -S(O) 2 OH.
  • Sulfonic ester may refer to -S(O) 2 OR, wherein R is a group such as an alkyl as in sulfonic alkyl ester.
  • “Thio” may refer to -S-.
  • Alkylthio may refer to RS- wherein a thiol radical is substituted with an alkyl radical R. Examples may include methylthio, ethylthio, butylthio, and the like.
  • Arylthio may refer to R'S-, wherein a thio radical is substituted with an aryl radical, as herein defined. Examples may include, but are not limited to, phenylthio, phenylthiomethyl, and the like.
  • Alkylthio sulfonic acid may refer to the radical HO 3 SRS-, wherein an alkylthioradical is substituted with a sulfonic acid radical.
  • Thiosulfenyl may refer to -S-SH.
  • Acyl may refer to a carbonyl or thiocarbonyl group bonded to a radical selected from, for example, hydrido, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, alkoxyalkyl, haloalkoxy, aryl, heterocyclyl, heteroaryl, alkylsulfmylalkyl, alkylsulfonylalkyl, aralkyl, cycloalkyl, cycloalkylalkyl, cycloalkenyl, alkylthio, arylthio, amino, alkylamino, dialkylamino, aralkoxy, arylthio, and alkylthioalkyl.
  • acyl may include formyl, acetyl, benzoyl, trifluoroacetyl, phthaloyl, malonyl, nicotinyl, and the like.
  • acylthiol and “acyldisulfide” may refer to the radicals RCOS- and RCOSS- respectively.
  • Carbonyloxy may refer to -OCOR.
  • Alkoxycarbonyl may refer to -COOR.
  • Carboxyl may refer to -COOH.
  • the following definitions apply to terms used to describe certain Effective Compounds discussed herein. In some embodiments, these definitions apply to Formulas Ia-Id, II, III, Ilia, V, VI, VII, VIII, IX and X:
  • Alkyl may refer to monovalent alkyl groups having from 1 to 10 carbon atoms, preferably from 1 to 5 carbon atoms and more preferably 1 to 3 carbon atoms. This term may be exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, n-pentyl and the like.
  • Substituted alkyl may refer to an alkyl group, of from 1 to 10 carbon atoms, preferably, 1 to 5 carbon atoms, having from 1 to 5 substituents, preferably 1 to 3 substituents, independently selected from the group consisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy, aryloxyaryl, substituted aryloxyaryl, cyano, halogen, hydroxyl, nitro, oxo, thioxo, carboxyl, carboxyl esters, cycloalkyl, substituted cycloalkyl, thiol, alkylthio, substituted alkylthio, arylthio, substituted arylthio, cycloalkylthio,
  • Alkoxy may refer to the group “alkyl-O— " which may include, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy and the like.
  • Substituted alkoxy may refer to the group “substituted alkyl-O—”.
  • Acyl may refer to the groups H-C(O)-, alkyl-C(O)-, substituted alkyl- C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, aryl-C(O)-, substituted aryl-C(O)-, heteroaryl-C(O)— , substituted heteroaryl-C(O), heterocyclic- C(O)-, and substituted hetero cyclic-C (O)- provided that a nitrogen atom of the heterocyclic or substituted heterocyclic is not bound to the -C(O)- group wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, substituted
  • aminoacyl or as a prefix “carbamoyl” or “carboxamide” or “substituted carbamoyl” or “substituted carboxamide” may refer to the group — C(O)NR 42 R 42 where each R 42 is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic and where each R 42 is joined to form together with the nitrogen atom a heterocyclic or substituted heterocyclic wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, hetero
  • Acyloxy may refer to the groups alkyl-C(O)O ⁇ , substituted alkyl-C(O)O ⁇ , alkenyl-C(O)O ⁇ , substituted alkenyl-C(O)O ⁇ , alkynyl-C(O)O ⁇ , substituted alkynyl- C(O)O-, aryl-C(O)O ⁇ , substituted aryl-C(O)O ⁇ , cycloalkyl-C(O)O ⁇ , substituted cycloalkyl-C(O)O ⁇ , heteroaryl-C(O)O ⁇ , substituted heteroaryl-C(O)O ⁇ , heterocyclic-C(O)O ⁇ , and substituted heterocyclic-C(O)O ⁇ wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl
  • Alkenyl may refer to alkenyl groups preferably having from 2 to 6 carbon atoms and more preferably 2 to 4 carbon atoms and having at least 1 and preferably from 1 to 2 sites of alkenyl unsaturation.
  • Substituted alkenyl may refer to alkenyl groups having from 1 to 3 substituents, and preferably 1 to 2 substituents, selected from the group consisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aryl, substituted aryl, aryloxy, substituted aryloxy, cyano, halogen, hydroxyl, nitro, carboxyl, carboxyl esters, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic.
  • Alkynyl may refer to alkynyl group preferably having from 2 to 6 carbon atoms and more preferably 2 to 3 carbon atoms and having at least 1 and preferably from 1-2 sites of alkynyl unsaturation.
  • Substituted alkynyl may refer to alkynyl groups having from 1 to 3 substituents, and preferably 1 to 2 substituents, selected from the group consisting of alkoxy, substituted alkoxy, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aryl, substituted aryl, aryloxy, substituted aryloxy, cyano, halogen, hydroxyl, nitro, carboxyl, carboxyl esters, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic.
  • Amino may refer to the group -NH 2 .
  • Substituted amino may refer to the group --NR 41 R 41 , where each R 41 group is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, ⁇ SO 2 -alkyl, ⁇ SO 2 -substituted alkyl, ⁇ SO 2 -alkenyl, -SO 2 - substituted alkenyl, ⁇ SO 2 -cycloalkyl, ⁇ SO 2 -substituted cycloalkyl, — SO 2 -aryl, -SO 2 - substituted aryl, — SO 2 -heteroaryl, — SO 2 -substituted heteroaryl, ⁇ SO 2 -heterocycl
  • Acylamino may refer to the groups ⁇ NR 45 C(O)alkyl, ⁇ NR 45 C(O)substituted alkyl, -NR 45 C(O)cycloalkyl, -NR 45 C(O)substituted cycloalkyl, -NR 45 C(O)alkenyl, - -NR 45 C(O)substituted alkenyl, -NR 45 C(O)alkynyl, -NR 45 C(O)substituted alkynyl, - NR 45 C(O)aryl, -NR 45 C(O)substituted aryl, -NR 45 C(O)heteroaryl, NR 45 C(O)substituted heteroaryl, -NR 45 C(O)heterocyclic, and ⁇ NR 45 C(O)substituted heterocyclic where R 45 is hydrogen or alkyl and wherein alkyl, substituted alkyl, alky
  • NR 46 C(O)O-substituted alkyl -NR 46 C(O)O-alkenyl, -NR 46 C(O)O-substituted alkenyl, -NR 46 C(O)O-alkynyl, -NR 46 C(O)O-substituted alkynyl, -NR 46 C(O)O- cycloalkyl, --NR 46 C(O)O-substituted cycloalkyl, -NR 46 C(O)O-aryl, -NR 46 C(O)O- substituted aryl, ⁇ NR 46 C(O)O-heteroaryl, ⁇ NR 46 C(O)O-substituted heteroaryl, - NR 46 C(O)O-heterocyclic, and ⁇ NR 46 C(O)O-substituted heterocyclic where R 46 is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,
  • Aminocarbonyloxy or as a prefix “carbamoyloxy” or “substituted carbamoyloxy” may refer to the groups --OC(O)NR 47 R 47 where each R 47 is independently hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic or where each R 47 is joined to form, together with the nitrogen atom a heterocyclic or substituted heterocyclic and wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic are as
  • Aminocarbonylamino may refer to the group --NR 49 C(O)NR 49 -- where R 49 is selected from the group consisting of hydrogen and alkyl.
  • Aryl or “Ar” may refer to a monovalent aromatic carbocyclic group of from
  • Preferred aryls may include phenyl and naphthyl.
  • Substituted aryl may refer to aryl groups, as defined herein, which are substituted with from 1 to 4, preferably 1-3, substituents selected from the group consisting of hydroxy, acyl, acylamino, carbonylaminothio, acyloxy, alkyl, substituted alkyl, alkoxy, substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, amidino, amino, substituted amino, aminoacyl, aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy, hetero aryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl, carboxyl esters cyano, thiol, alkylthio, substituted alkylthio, aryl
  • Aryloxy may refer to the group aryl-O— that may include, by way of example, phenoxy, naphthoxy, and the like.
  • Substituted aryloxy may refer to substituted aryl-O— groups.
  • Aryloxyaryl may refer to the group -aryl-O-aryl.
  • Substituted aryloxyaryl may refer to aryloxyaryl groups substituted with from 1 to 3 substituents on either or both aryl rings as defined above for substituted aryl.
  • Carboxyl may refer to — COOH or salts thereof.
  • Carboxyl esters may refer to the groups -C(O)O-alkyl, — C(O)O-substituted alkyl, — C(O)O-aryl, and — C(O)O-substituted aryl wherein alkyl, substituted alkyl, aryl and substituted aryl are as defined herein.
  • Cycloalkyl may refer to cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple cyclic rings that may include, by way of example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl and the like.
  • Cycloalkoxy may refer to --O-cycloalkyl groups.
  • Substituted cycloalkoxy may refer to — O-substituted cycloalkyl groups.
  • Halo or “halogen” may refer to fluoro, chloro, bromo and iodo and preferably is fluoro or chloro.
  • Heteroaryl may refer to an aromatic group of from 1 to 15 carbon atoms, preferably from 1 to 10 carbon atoms, and 1 to 4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur within the ring.
  • Such heteroaryl groups can have a single ring (e.g., pyridinyl or furyl) or multiple condensed rings (e.g., indolizinyl or benzothienyl).
  • Preferred heteroaryls may include pyridinyl, pyrrolyl, indolyl, thiophenyl, and furyl.
  • “Substituted heteroaryl” may refer to heteroaryl groups that are substituted with from 1 to 3 substituents selected from the same group of substituents defined for substituted aryl.
  • Heteroaryloxy may refer to the group --O-heteroaryl and "substituted heteroaryloxy” may refer to the group --O-substituted heteroaryl.
  • Heterocycle or “heterocyclic” may refer to a saturated or unsaturated group having a single ring or multiple condensed rings, from 1 to 10 carbon atoms and from 1 to 4 heteroatoms selected from the group consisting of nitrogen, sulfur or oxygen within the ring wherein, in fused ring systems, one or more the rings can be aryl or heteroaryl provided that the point of attachment is at the heterocycle.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Zoology (AREA)
  • Dentistry (AREA)
  • Inorganic Chemistry (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Biomedical Technology (AREA)
  • Cardiology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Urology & Nephrology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Pulmonology (AREA)
  • Rheumatology (AREA)
  • Pain & Pain Management (AREA)
  • Psychiatry (AREA)
  • Hospice & Palliative Care (AREA)
  • Vascular Medicine (AREA)
  • Surgery (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne de manière générale la modulation d'un facteur inductible par l'hypoxie (HIF) au moyen des composés et des procédés décrits dans la présente invention. Ces composés et ces procédés peuvent être appliqués à la prévention, au prétraitement et/ou au traitement de conditions ou d'états associés à HIF, tel que des conditions liées à une hypoxie et à une ischémie ainsi que l'induction d'une stase.
PCT/US2007/079948 2006-09-28 2007-09-28 Procédés, compositions et articles de fabrication de composés modulant le hif Ceased WO2008040002A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US82733706P 2006-09-28 2006-09-28
US60/827,337 2006-09-28

Publications (3)

Publication Number Publication Date
WO2008040002A2 true WO2008040002A2 (fr) 2008-04-03
WO2008040002A9 WO2008040002A9 (fr) 2009-04-16
WO2008040002A3 WO2008040002A3 (fr) 2009-07-09

Family

ID=39032331

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2007/079948 Ceased WO2008040002A2 (fr) 2006-09-28 2007-09-28 Procédés, compositions et articles de fabrication de composés modulant le hif

Country Status (2)

Country Link
US (2) US20090011051A1 (fr)
WO (1) WO2008040002A2 (fr)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7569726B2 (en) 2007-04-18 2009-08-04 Amgen Inc. Indanone derivatives that inhibit prolyl hydroxylase
US7635715B2 (en) 2006-12-18 2009-12-22 Amgen Inc. Naphthalenone compounds exhibiting prolyl hydroxylase inhibitory activity, compositions, and uses thereof
US8030346B2 (en) 2007-05-04 2011-10-04 Amgen Inc. Heterocyclic quinolone derivatives that inhibit prolyl hydroxylase activity
US8048892B2 (en) 2006-12-18 2011-11-01 Amgen Inc. Azaquinolone based compounds exhibiting prolyl hydroxylase inhibitory activity, compositions, and uses thereof
US8048894B2 (en) 2007-04-18 2011-11-01 Amgen Inc. Quinolones and azaquinolones that inhibit prolyl hydroxylase
US8097620B2 (en) 2007-05-04 2012-01-17 Amgen Inc. Diazaquinolones that inhibit prolyl hydroxylase activity
US8158339B2 (en) 2008-07-07 2012-04-17 Rich Products Corporation Method of preserving a platelet concentrate under elevated xenon concentration and pressure with refrigeration
US8324208B2 (en) 2006-06-23 2012-12-04 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
USRE44613E1 (en) 2007-01-12 2013-11-26 Glaxosmithkline Llc N-substituted glycine derivatives: hydroxylase inhibitors
WO2013177478A3 (fr) * 2012-05-24 2014-01-16 Glaxosmithkline Intellectual Property (No.2) Limited Méthode de traitement
CN106146395A (zh) * 2015-03-27 2016-11-23 沈阳三生制药有限责任公司 3-羟基吡啶化合物、其制备方法及其制药用途
WO2017059427A1 (fr) * 2015-10-02 2017-04-06 Children's National Medical Center Procédés de contrôle et de détermination du pronostic d'avc, de maladie vasculaire périphérique, de choc et de drépanocytose et de ses complications
CN107417605A (zh) * 2017-08-02 2017-12-01 江苏艾立康药业股份有限公司 作用于脯氨酰羟化酶的吡啶衍生化合物
US10201570B2 (en) 2014-02-10 2019-02-12 Fred Hutchinson Cancer Research Center Halogen treatment of heart attack and ischemic injury
US11166451B2 (en) 2011-09-26 2021-11-09 Rich Technologies Holding Company, Llc Method for living tissue preservation
WO2022036188A1 (fr) * 2020-08-14 2022-02-17 Akebia Therapeutics, Inc. Composés inhibiteurs de phd, compositions et procédés d'utilisation

Families Citing this family (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6855510B2 (en) * 2001-03-20 2005-02-15 Dana Farber Cancer Institute, Inc. Pharmaceuticals and methods for treating hypoxia and screening methods therefor
WO2010005968A1 (fr) * 2008-07-07 2010-01-14 Rich Products Corporation Procédé pour le stockage de crustacés vivants
KR100976548B1 (ko) * 2008-09-12 2010-08-17 김희구 약재 성분의 방출이 조절되는 생약패드 및 이의 제조방법
US8551454B2 (en) * 2009-03-13 2013-10-08 Luitpold Pharmaceuticals, Inc. Device for intranasal administration
US8277781B2 (en) * 2009-03-13 2012-10-02 Luitpold Pharmaceuticals, Inc. Device for intranasal administration
US10980752B2 (en) 2009-09-14 2021-04-20 Bm Biotechnology Co., Ltd. Device for herbal medicine in which release of medicinal ingredient can be controlled, and manufacturing method thereof
WO2012037212A1 (fr) * 2010-09-14 2012-03-22 Dana-Farber Cancer Institute, Inc. Inhibiteurs de prolyl hydroxylase à titre d'agents d'atténuation du rayonnement et de protection contre le rayonnement
US9820981B2 (en) 2013-12-19 2017-11-21 The Cleveland Clinic Foundation Treatment of retinopathy of prematurity (ROP)
JP6822953B2 (ja) * 2014-11-25 2021-01-27 アプライド マテリアルズ インコーポレイテッドＡｐｐｌｉｅｄ Ｍａｔｅｒｉａｌｓ，Ｉｎｃｏｒｐｏｒａｔｅｄ 基板キャリア及びパージチャンバの環境制御を伴う基板処理のシステム、装置、及び方法
EP4512819A3 (fr) 2015-08-12 2025-08-06 The General Hospital Corporation Compositions et procédés qui favorisent la réponse à l'hypoxie ou à l'hypoxie pour le traitement et la prévention d'un dysfonctionnement mitochondrial et de troubles du stress oxydatif
EP3383415B1 (fr) 2015-12-01 2021-03-31 Cornell University Utilisation de chélateurs du fer mitochondrial pour le traitement de la bronchopneumopathie chronique obstructive
JP7386162B2 (ja) * 2017-12-20 2023-11-24 フィジーン、エルエルシー 線維芽細胞の再生活性の増強
WO2019173283A1 (fr) 2018-03-05 2019-09-12 Marquette University Procédé et appareil de prédiction du niveau d'hémoglobine non invasifs
US20210244700A1 (en) * 2018-05-15 2021-08-12 Duke University Compositions and methods for preventing and/or reducing ischemia after surgical incisions
US20210113524A1 (en) * 2018-05-31 2021-04-22 The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. Methods of treating myeloproliferative disorders
US12544403B2 (en) 2019-01-10 2026-02-10 The General Hospital Corporation Methods to treat mitochondrial-associated dysfunctions or diseases
CN111001016B (zh) * 2019-12-20 2022-06-17 中国人民解放军陆军军医大学 一种vha抑制dr5在缺氧下精子生成减少的检测方法
CN112618697B (zh) * 2020-12-22 2021-10-01 浙江师范大学 一种水产低温保护剂及其用途
CN119700755B (zh) * 2024-02-05 2026-01-30 深圳信立泰药业股份有限公司 化合物在制备预防和/或治疗高原病的药物中的应用
CN118084903A (zh) * 2024-02-27 2024-05-28 贵州医科大学 一种β-咔伯啉硫化氢供体衍生物及其制备方法与应用

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK2298301T3 (en) * 2001-12-06 2018-10-01 Fibrogen Inc MEDICINALS FOR TREATMENT OF ANEMIA ASSOCIATED WITH Kidney Disease
WO2004108681A1 (fr) * 2003-06-06 2004-12-16 Fibrogen, Inc. Composes heteroaryliques contenant de l'azote et leur utilisation dans l'augmentation de l'erythropoietine endogene
CN1901795B (zh) * 2003-10-22 2014-03-26 弗雷德哈钦森癌症研究中心 用于在细胞，组织，器官，和有机体中诱导停滞的方法，组合物和装置
US20050136125A1 (en) * 2003-10-22 2005-06-23 Roth Mark B. Methods, compositions and devices for inducing stasis in cells, tissues, organs, and organisms
US20050170019A1 (en) * 2003-10-22 2005-08-04 Fred Hutchinson Cancer Research Center Methods, compositions and devices for inducing stasis in cells
CA2605631A1 (fr) * 2005-04-20 2006-10-26 Fred Hutchinson Cancer Research Center Procedes, compositions et articles manufactures permettant d'ameliorer la capacite de survie de cellules, de tissus, d'organes et d'organismes
CA2646674A1 (fr) * 2006-04-20 2007-11-01 Fred Hutchinson Cancer Research Center Procedes, compositions et articles fabriques destines au traitement du choc et d'autres etats defavorables

Cited By (28)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11643397B2 (en) 2006-06-23 2023-05-09 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
US8324208B2 (en) 2006-06-23 2012-12-04 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
US10336711B2 (en) 2006-06-23 2019-07-02 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
US8815884B2 (en) 2006-06-23 2014-08-26 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
US10035779B2 (en) 2006-06-23 2018-07-31 GlaxoSmithKline, LLC Prolyl hydroxylase inhibitors
US8557834B2 (en) 2006-06-23 2013-10-15 Glaxosmithkline Llc Prolyl hydroxylase inhibitors
US8048892B2 (en) 2006-12-18 2011-11-01 Amgen Inc. Azaquinolone based compounds exhibiting prolyl hydroxylase inhibitory activity, compositions, and uses thereof
US7635715B2 (en) 2006-12-18 2009-12-22 Amgen Inc. Naphthalenone compounds exhibiting prolyl hydroxylase inhibitory activity, compositions, and uses thereof
US7928139B2 (en) 2006-12-18 2011-04-19 Amgen Inc. Naphthalenone compounds exhibiting prolyl hydroxylase inhibitory activity, compositions, and uses thereof
USRE44613E1 (en) 2007-01-12 2013-11-26 Glaxosmithkline Llc N-substituted glycine derivatives: hydroxylase inhibitors
US8349868B2 (en) 2007-04-18 2013-01-08 Amgen Inc. Azaquinolones that inhibit prolyl hydroxylase
US8048894B2 (en) 2007-04-18 2011-11-01 Amgen Inc. Quinolones and azaquinolones that inhibit prolyl hydroxylase
US7569726B2 (en) 2007-04-18 2009-08-04 Amgen Inc. Indanone derivatives that inhibit prolyl hydroxylase
US8097620B2 (en) 2007-05-04 2012-01-17 Amgen Inc. Diazaquinolones that inhibit prolyl hydroxylase activity
US8030346B2 (en) 2007-05-04 2011-10-04 Amgen Inc. Heterocyclic quinolone derivatives that inhibit prolyl hydroxylase activity
US8158339B2 (en) 2008-07-07 2012-04-17 Rich Products Corporation Method of preserving a platelet concentrate under elevated xenon concentration and pressure with refrigeration
US11166451B2 (en) 2011-09-26 2021-11-09 Rich Technologies Holding Company, Llc Method for living tissue preservation
WO2013177478A3 (fr) * 2012-05-24 2014-01-16 Glaxosmithkline Intellectual Property (No.2) Limited Méthode de traitement
US10201570B2 (en) 2014-02-10 2019-02-12 Fred Hutchinson Cancer Research Center Halogen treatment of heart attack and ischemic injury
US12016880B2 (en) 2014-02-10 2024-06-25 Fred Hutchinson Cancer Center Halogen treatment of heart attack and ischemic injury
CN106146395B (zh) * 2015-03-27 2019-01-01 沈阳三生制药有限责任公司 3-羟基吡啶化合物、其制备方法及其制药用途
CN106146395A (zh) * 2015-03-27 2016-11-23 沈阳三生制药有限责任公司 3-羟基吡啶化合物、其制备方法及其制药用途
WO2017059427A1 (fr) * 2015-10-02 2017-04-06 Children's National Medical Center Procédés de contrôle et de détermination du pronostic d'avc, de maladie vasculaire périphérique, de choc et de drépanocytose et de ses complications
CN107417605A (zh) * 2017-08-02 2017-12-01 江苏艾立康药业股份有限公司 作用于脯氨酰羟化酶的吡啶衍生化合物
WO2022036188A1 (fr) * 2020-08-14 2022-02-17 Akebia Therapeutics, Inc. Composés inhibiteurs de phd, compositions et procédés d'utilisation
CN116670131A (zh) * 2020-08-14 2023-08-29 阿克比治疗有限公司 Phd抑制剂化合物、组合物和使用方法
JP2023537611A (ja) * 2020-08-14 2023-09-04 アケビア セラピューティクス インコーポレイテッド Ｐｈｄ阻害剤化合物、組成物、及び使用方法
JP7832179B2 (ja) 2020-08-14 2026-03-17 アケビア セラピューティクス インコーポレイテッド Ｐｈｄ阻害剤化合物、組成物、及び使用方法

Also Published As

Publication number Publication date
US20090011051A1 (en) 2009-01-08
WO2008040002A9 (fr) 2009-04-16
WO2008040002A3 (fr) 2009-07-09
US20120282353A1 (en) 2012-11-08

Similar Documents

Publication Publication Date Title
US20090011051A1 (en) Methods, Compositions and Articles of Manufacture for HIF Modulating Compounds
US20080226750A1 (en) Methods, Compositions and Articles of Manufacture for Treating Shock and Other Adverse Conditions
US20120040024A1 (en) Methods and compositions for enhancing survivability of cells, tissues, organs and organisms
US7238469B2 (en) Carbon monoxide improves outcomes in tissue and organ transplants and suppresses apoptosis
Bos et al. Hydrogen sulfide: physiological properties and therapeutic potential in ischaemia
US20080318864A1 (en) Methods and compositions regarding polychalcogenide compositions
CN105579045A (zh) 用于器官停滞、保护和保存以及减少组织损伤的方法
AU2006254897A1 (en) Improved treatment for anemia using a HIF-alpha stabilising agent
JP2007528855A (ja) 亜硝酸塩によって特定の心臓血管状態を処置する方法
AU2002318377A1 (en) Carbon monoxide improves outcomes in tissue and organ transplants and suppresses apoptosis
AU2007254300A1 (en) Methods of treating acute blood loss anemia with a cross-linked hemoglobin blood substitute
Dosier et al. Antagonists of the system L neutral amino acid transporter (LAT) promote endothelial adhesivity of human red blood cells
CN101203231A (zh) 增强细胞、组织、器官和生物的存活力的方法、组合物和制造品
Nakamura et al. Heme oxygenase does not contribute to control of basal vascular tone in isolated blood-perfused rat lung
RU2544958C2 (ru) СПОСОБ ЗАЩИТЫ КЛЕТОК ОТ ПОВРЕЖДЕНИЯ ПРИ ГИПОКСИИ ПУТЕМ ГЛУТАТИОНИЛИРОВАНИЯ КАТАЛИТИЧЕСКОЙ СУБЪЕДИНИЦЫ Na, К-АТФазы
Kougia et al. Pharmaceutical Interventions for the Management of Hemorrhagic Shock in Hepatic Surgery: An Experimental Swine Model Outcome

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07843518

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 07843518

Country of ref document: EP

Kind code of ref document: A2